Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric

preprint OA: closed
Full text JSON View at publisher

Abstract

Background: The phylogenetic bootstrap, first proposed by Felsenstein in 1985, is a critically important statistical method in assessing the robusticity of phylogenetic datasets. Core to its concept was the use of pseudo sampling - assessing the data by generating new replicates derived from the initial dataset that was used to generate the phylogeny. In this way, phylogenetic support metrics could overcome the lack of perfect, infinite data. With infinite data, however, it is possible to sample smaller replicates directly from the data to obtain both the phylogeny and its statistical robusticity in the same analysis. Due to the growth of whole genome sequencing, the depth and breadth of our datasets have greatly expanded and are set to only expand further. With genome-scale datasets comprising thousands of genes, we can now obtain a proxy for infinite data. Accordingly, we can potentially abandon the notion of pseudo sampling and instead randomly sample small subsets of genes from the thousands of genes in our analyses. Methods: We introduce Scoutknife, a jackknife-style subsampling implementation that generates 100 datasets by randomly sampling a small number of genes from an initial large-gene dataset to jointly establish both a phylogenetic hypothesis and assess its robusticity. We assess its effectiveness by using 18 previously published datasets and 100 simulation studies. Results: We show that Scoutknife is conservative and informative as to conflicts and incongruence across the whole genome, without the need for subsampling based on traditional model selection criteria. Conclusions: Scoutknife reliably achieves comparable results to selecting the best genes on both real and simulation datasets, while being resistant to the potential biases caused by selecting for model fit. As the amount of genome data grows, it becomes an even more exciting option to assess the robusticity of phylogenetic hypotheses.
Full text 183,411 characters · extracted from preprint-html · click to expand
Scoutknife: A naïve, whole genome informed... | F1000Research "use strict";function _typeof(t){return(_typeof="function"==typeof Symbol&&"symbol"==typeof Symbol.iterator?function(t){return typeof t}:function(t){return t&&"function"==typeof Symbol&&t.constructor===Symbol&&t!==Symbol.prototype?"symbol":typeof t})(t)}!function(){var t=function(){var t,e,o=[],n=window,r=n;for(;r;){try{if(r.frames.__tcfapiLocator){t=r;break}}catch(t){}if(r===n.top)break;r=r.parent}t||(!function t(){var e=n.document,o=!!n.frames.__tcfapiLocator;if(!o)if(e.body){var r=e.createElement("iframe");r.style.cssText="display:none",r.name="__tcfapiLocator",e.body.appendChild(r)}else setTimeout(t,5);return!o}(),n.__tcfapi=function(){for(var t=arguments.length,n=new Array(t),r=0;r 3&&2===parseInt(n[1],10)&&"boolean"==typeof n[3]&&(e=n[3],"function"==typeof n[2]&&n[2]("set",!0)):"ping"===n[0]?"function"==typeof n[2]&&n[2]({gdprApplies:e,cmpLoaded:!1,cmpStatus:"stub"}):o.push(n)},n.addEventListener("message",(function(t){var e="string"==typeof t.data,o={};if(e)try{o=JSON.parse(t.data)}catch(t){}else o=t.data;var n="object"===_typeof(o)&&null!==o?o.__tcfapiCall:null;n&&window.__tcfapi(n.command,n.version,(function(o,r){var a={__tcfapiReturn:{returnValue:o,success:r,callId:n.callId}};t&&t.source&&t.source.postMessage&&t.source.postMessage(e?JSON.stringify(a):a,"*")}),n.parameter)}),!1))};"undefined"!=typeof module?module.exports=t:t()}(); dataLayer = dataLayer || []; // Standard GTM initialization - Google Consent Mode handles consent automatically (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start': new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0], j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src= 'https://www.googletagmanager.com/gtm.js?id='+i+dl+ '>m_auth=hzk0Vc3qFsQYhCrIoHz68A>m_preview=env-1>m_cookies_win=x';f.parentNode.insertBefore(j,f); })(window,document,'script','dataLayer','GTM-MWFK8L5J'); ;window.NREUM||(NREUM={});NREUM.init={distributed_tracing:{enabled:true},privacy:{cookies_enabled:true},ajax:{deny_list:["bam.nr-data.net"]}}; ;NREUM.loader_config={accountID:"438030",trustKey:"438030",agentID:"772317073",licenseKey:"97f8f67f26",applicationID:"772317073"} ;NREUM.info={beacon:"bam.nr-data.net",errorBeacon:"bam.nr-data.net",licenseKey:"97f8f67f26",applicationID:"772317073",sa:1} ;/*! For license information please see nr-loader-spa-1.236.0.min.js.LICENSE.txt */ (()=>{"use strict";var e,t,r={5763:(e,t,r)=>{r.d(t,{P_:()=>l,Mt:()=>g,C5:()=>s,DL:()=>v,OP:()=>T,lF:()=>D,Yu:()=>y,Dg:()=>h,CX:()=>c,GE:()=>b,sU:()=>_});var n=r(8632),i=r(9567);const o={beacon:n.ce.beacon,errorBeacon:n.ce.errorBeacon,licenseKey:void 0,applicationID:void 0,sa:void 0,queueTime:void 0,applicationTime:void 0,ttGuid:void 0,user:void 0,account:void 0,product:void 0,extra:void 0,jsAttributes:{},userAttributes:void 0,atts:void 0,transactionName:void 0,tNamePlain:void 0},a={};function s(e){if(!e)throw new Error("All info objects require an agent identifier!");if(!a[e])throw new Error("Info for ".concat(e," was never set"));return a[e]}function c(e,t){if(!e)throw new Error("All info objects require an agent identifier!");a[e]=(0,i.D)(t,o),(0,n.Qy)(e,a[e],"info")}var u=r(7056);const d=()=>{const e={blockSelector:"[data-nr-block]",maskInputOptions:{password:!0}};return{allow_bfcache:!0,privacy:{cookies_enabled:!0},ajax:{deny_list:void 0,enabled:!0,harvestTimeSeconds:10},distributed_tracing:{enabled:void 0,exclude_newrelic_header:void 0,cors_use_newrelic_header:void 0,cors_use_tracecontext_headers:void 0,allowed_origins:void 0},session:{domain:void 0,expiresMs:u.oD,inactiveMs:u.Hb},ssl:void 0,obfuscate:void 0,jserrors:{enabled:!0,harvestTimeSeconds:10},metrics:{enabled:!0},page_action:{enabled:!0,harvestTimeSeconds:30},page_view_event:{enabled:!0},page_view_timing:{enabled:!0,harvestTimeSeconds:30,long_task:!1},session_trace:{enabled:!0,harvestTimeSeconds:10},harvest:{tooManyRequestsDelay:60},session_replay:{enabled:!1,harvestTimeSeconds:60,sampleRate:.1,errorSampleRate:.1,maskTextSelector:"*",maskAllInputs:!0,get blockClass(){return"nr-block"},get ignoreClass(){return"nr-ignore"},get maskTextClass(){return"nr-mask"},get blockSelector(){return e.blockSelector},set blockSelector(t){e.blockSelector+=",".concat(t)},get maskInputOptions(){return e.maskInputOptions},set maskInputOptions(t){e.maskInputOptions={...t,password:!0}}},spa:{enabled:!0,harvestTimeSeconds:10}}},f={};function l(e){if(!e)throw new Error("All configuration objects require an agent identifier!");if(!f[e])throw new Error("Configuration for ".concat(e," was never set"));return f[e]}function h(e,t){if(!e)throw new Error("All configuration objects require an agent identifier!");f[e]=(0,i.D)(t,d()),(0,n.Qy)(e,f[e],"config")}function g(e,t){if(!e)throw new Error("All configuration objects require an agent identifier!");var r=l(e);if(r){for(var n=t.split("."),i=0;i {r.d(t,{D:()=>i});var n=r(50);function i(e,t){try{if(!e||"object"!=typeof e)return(0,n.Z)("Setting a Configurable requires an object as input");if(!t||"object"!=typeof t)return(0,n.Z)("Setting a Configurable requires a model to set its initial properties");const r=Object.create(Object.getPrototypeOf(t),Object.getOwnPropertyDescriptors(t)),o=0===Object.keys(r).length?e:r;for(let a in o)if(void 0!==e[a])try{"object"==typeof e[a]&&"object"==typeof t[a]?r[a]=i(e[a],t[a]):r[a]=e[a]}catch(e){(0,n.Z)("An error occurred while setting a property of a Configurable",e)}return r}catch(e){(0,n.Z)("An error occured while setting a Configurable",e)}}},6818:(e,t,r)=>{r.d(t,{Re:()=>i,gF:()=>o,q4:()=>n});const n="1.236.0",i="PROD",o="CDN"},385:(e,t,r)=>{r.d(t,{FN:()=>a,IF:()=>u,Nk:()=>f,Tt:()=>s,_A:()=>o,il:()=>n,pL:()=>c,v6:()=>i,w1:()=>d});const n="undefined"!=typeof window&&!!window.document,i="undefined"!=typeof WorkerGlobalScope&&("undefined"!=typeof self&&self instanceof WorkerGlobalScope&&self.navigator instanceof WorkerNavigator||"undefined"!=typeof globalThis&&globalThis instanceof WorkerGlobalScope&&globalThis.navigator instanceof WorkerNavigator),o=n?window:"undefined"!=typeof WorkerGlobalScope&&("undefined"!=typeof self&&self instanceof WorkerGlobalScope&&self||"undefined"!=typeof globalThis&&globalThis instanceof WorkerGlobalScope&&globalThis),a=""+o?.location,s=/iPad|iPhone|iPod/.test(navigator.userAgent),c=s&&"undefined"==typeof SharedWorker,u=(()=>{const e=navigator.userAgent.match(/Firefox[/\s](\d+\.\d+)/);return Array.isArray(e)&&e.length>=2?+e[1]:0})(),d=Boolean(n&&window.document.documentMode),f=!!navigator.sendBeacon},1117:(e,t,r)=>{r.d(t,{w:()=>o});var n=r(50);const i={agentIdentifier:"",ee:void 0};class o{constructor(e){try{if("object"!=typeof e)return(0,n.Z)("shared context requires an object as input");this.sharedContext={},Object.assign(this.sharedContext,i),Object.entries(e).forEach((e=>{let[t,r]=e;Object.keys(i).includes(t)&&(this.sharedContext[t]=r)}))}catch(e){(0,n.Z)("An error occured while setting SharedContext",e)}}}},8e3:(e,t,r)=>{r.d(t,{L:()=>d,R:()=>c});var n=r(2177),i=r(1284),o=r(4322),a=r(3325);const s={};function c(e,t){const r={staged:!1,priority:a.p[t]||0};u(e),s[e].get(t)||s[e].set(t,r)}function u(e){e&&(s[e]||(s[e]=new Map))}function d(){let e=arguments.length>0&&void 0!==arguments[0]?arguments[0]:"",t=arguments.length>1&&void 0!==arguments[1]?arguments[1]:"feature";if(u(e),!e||!s[e].get(t))return a(t);s[e].get(t).staged=!0;const r=[...s[e]];function a(t){const r=e?n.ee.get(e):n.ee,a=o.X.handlers;if(r.backlog&&a){var s=r.backlog[t],c=a[t];if(c){for(var u=0;s&&u {let[t,r]=e;return r.staged}))&&(r.sort(((e,t)=>e[1].priority-t[1].priority)),r.forEach((e=>{let[t]=e;a(t)})))}function f(e,t){var r=e[1];(0,i.D)(t[r],(function(t,r){var n=e[0];if(r[0]===n){var i=r[1],o=e[3],a=e[2];i.apply(o,a)}}))}},2177:(e,t,r)=>{r.d(t,{c:()=>f,ee:()=>u});var n=r(8632),i=r(2210),o=r(1284),a=r(5763),s="nr@context";let c=(0,n.fP)();var u;function d(){}function f(e){return(0,i.X)(e,s,l)}function l(){return new d}function h(){u.aborted=!0,u.backlog={}}c.ee?u=c.ee:(u=function e(t,r){var n={},c={},f={},g=!1;try{g=16===r.length&&(0,a.OP)(r).isolatedBacklog}catch(e){}var p={on:b,addEventListener:b,removeEventListener:y,emit:v,get:x,listeners:w,context:m,buffer:A,abort:h,aborted:!1,isBuffering:E,debugId:r,backlog:g?{}:t&&"object"==typeof t.backlog?t.backlog:{}};return p;function m(e){return e&&e instanceof d?e:e?(0,i.X)(e,s,l):l()}function v(e,r,n,i,o){if(!1!==o&&(o=!0),!u.aborted||i){t&&o&&t.emit(e,r,n);for(var a=m(n),s=w(e),d=s.length,f=0;fn,p:()=>i});var n=r(2177).ee.get("handle");function i(e,t,r,i,o){o?(o.buffer([e],i),o.emit(e,t,r)):(n.buffer([e],i),n.emit(e,t,r))}},4322:(e,t,r)=>{r.d(t,{X:()=>o});var n=r(5546);o.on=a;var i=o.handlers={};function o(e,t,r,o){a(o||n.E,i,e,t,r)}function a(e,t,r,i,o){o||(o="feature"),e||(e=n.E);var a=t[o]=t[o]||{};(a[r]=a[r]||[]).push([e,i])}},3239:(e,t,r)=>{r.d(t,{bP:()=>s,iz:()=>c,m$:()=>a});var n=r(385);let i=!1,o=!1;try{const e={get passive(){return i=!0,!1},get signal(){return o=!0,!1}};n._A.addEventListener("test",null,e),n._A.removeEventListener("test",null,e)}catch(e){}function a(e,t){return i||o?{capture:!!e,passive:i,signal:t}:!!e}function s(e,t){let r=arguments.length>2&&void 0!==arguments[2]&&arguments[2],n=arguments.length>3?arguments[3]:void 0;window.addEventListener(e,t,a(r,n))}function c(e,t){let r=arguments.length>2&&void 0!==arguments[2]&&arguments[2],n=arguments.length>3?arguments[3]:void 0;document.addEventListener(e,t,a(r,n))}},4402:(e,t,r)=>{r.d(t,{Ht:()=>u,M:()=>c,Rl:()=>a,ky:()=>s});var n=r(385);const i="xxxxxxxx-xxxx-4xxx-yxxx-xxxxxxxxxxxx";function o(e,t){return e?15&e[t]:16*Math.random()|0}function a(){const e=n._A?.crypto||n._A?.msCrypto;let t,r=0;return e&&e.getRandomValues&&(t=e.getRandomValues(new Uint8Array(31))),i.split("").map((e=>"x"===e?o(t,++r).toString(16):"y"===e?(3&o()|8).toString(16):e)).join("")}function s(e){const t=n._A?.crypto||n._A?.msCrypto;let r,i=0;t&&t.getRandomValues&&(r=t.getRandomValues(new Uint8Array(31)));const a=[];for(var s=0;s {r.d(t,{Bq:()=>n,Hb:()=>o,oD:()=>i});const n="NRBA",i=144e5,o=18e5},7894:(e,t,r)=>{function n(){return Math.round(performance.now())}r.d(t,{z:()=>n})},7243:(e,t,r)=>{r.d(t,{e:()=>o});var n=r(385),i={};function o(e){if(e in i)return i[e];if(0===(e||"").indexOf("data:"))return{protocol:"data"};let t;var r=n._A?.location,o={};if(n.il)t=document.createElement("a"),t.href=e;else try{t=new URL(e,r.href)}catch(e){return o}o.port=t.port;var a=t.href.split("://");!o.port&&a[1]&&(o.port=a[1].split("/")[0].split("@").pop().split(":")[1]),o.port&&"0"!==o.port||(o.port="https"===a[0]?"443":"80"),o.hostname=t.hostname||r.hostname,o.pathname=t.pathname,o.protocol=a[0],"/"!==o.pathname.charAt(0)&&(o.pathname="/"+o.pathname);var s=!t.protocol||":"===t.protocol||t.protocol===r.protocol,c=t.hostname===r.hostname&&t.port===r.port;return o.sameOrigin=s&&(!t.hostname||c),"/"===o.pathname&&(i[e]=o),o}},50:(e,t,r)=>{function n(e,t){"function"==typeof console.warn&&(console.warn("New Relic: ".concat(e)),t&&console.warn(t))}r.d(t,{Z:()=>n})},2587:(e,t,r)=>{r.d(t,{N:()=>c,T:()=>u});var n=r(2177),i=r(5546),o=r(8e3),a=r(3325);const s={stn:[a.D.sessionTrace],err:[a.D.jserrors,a.D.metrics],ins:[a.D.pageAction],spa:[a.D.spa],sr:[a.D.sessionReplay,a.D.sessionTrace]};function c(e,t){const r=n.ee.get(t);e&&"object"==typeof e&&(Object.entries(e).forEach((e=>{let[t,n]=e;void 0===u[t]&&(s[t]?s[t].forEach((e=>{n?(0,i.p)("feat-"+t,[],void 0,e,r):(0,i.p)("block-"+t,[],void 0,e,r),(0,i.p)("rumresp-"+t,[Boolean(n)],void 0,e,r)})):n&&(0,i.p)("feat-"+t,[],void 0,void 0,r),u[t]=Boolean(n))})),Object.keys(s).forEach((e=>{void 0===u[e]&&(s[e]?.forEach((t=>(0,i.p)("rumresp-"+e,[!1],void 0,t,r))),u[e]=!1)})),(0,o.L)(t,a.D.pageViewEvent))}const u={}},2210:(e,t,r)=>{r.d(t,{X:()=>i});var n=Object.prototype.hasOwnProperty;function i(e,t,r){if(n.call(e,t))return e[t];var i=r();if(Object.defineProperty&&Object.keys)try{return Object.defineProperty(e,t,{value:i,writable:!0,enumerable:!1}),i}catch(e){}return e[t]=i,i}},1284:(e,t,r)=>{r.d(t,{D:()=>n});const n=(e,t)=>Object.entries(e||{}).map((e=>{let[r,n]=e;return t(r,n)}))},4351:(e,t,r)=>{r.d(t,{P:()=>o});var n=r(2177);const i=()=>{const e=new WeakSet;return(t,r)=>{if("object"==typeof r&&null!==r){if(e.has(r))return;e.add(r)}return r}};function o(e){try{return JSON.stringify(e,i())}catch(e){try{n.ee.emit("internal-error",[e])}catch(e){}}}},3960:(e,t,r)=>{r.d(t,{K:()=>a,b:()=>o});var n=r(3239);function i(){return"undefined"==typeof document||"complete"===document.readyState}function o(e,t){if(i())return e();(0,n.bP)("load",e,t)}function a(e){if(i())return e();(0,n.iz)("DOMContentLoaded",e)}},8632:(e,t,r)=>{r.d(t,{EZ:()=>u,Qy:()=>c,ce:()=>o,fP:()=>a,gG:()=>d,mF:()=>s});var n=r(7894),i=r(385);const o={beacon:"bam.nr-data.net",errorBeacon:"bam.nr-data.net"};function a(){return i._A.NREUM||(i._A.NREUM={}),void 0===i._A.newrelic&&(i._A.newrelic=i._A.NREUM),i._A.NREUM}function s(){let e=a();return e.o||(e.o={ST:i._A.setTimeout,SI:i._A.setImmediate,CT:i._A.clearTimeout,XHR:i._A.XMLHttpRequest,REQ:i._A.Request,EV:i._A.Event,PR:i._A.Promise,MO:i._A.MutationObserver,FETCH:i._A.fetch}),e}function c(e,t,r){let i=a();const o=i.initializedAgents||{},s=o[e]||{};return Object.keys(s).length||(s.initializedAt={ms:(0,n.z)(),date:new Date}),i.initializedAgents={...o,[e]:{...s,[r]:t}},i}function u(e,t){a()[e]=t}function d(){return function(){let e=a();const t=e.info||{};e.info={beacon:o.beacon,errorBeacon:o.errorBeacon,...t}}(),function(){let e=a();const t=e.init||{};e.init={...t}}(),s(),function(){let e=a();const t=e.loader_config||{};e.loader_config={...t}}(),a()}},7956:(e,t,r)=>{r.d(t,{N:()=>i});var n=r(3239);function i(e){let t=arguments.length>1&&void 0!==arguments[1]&&arguments[1],r=arguments.length>2?arguments[2]:void 0,i=arguments.length>3?arguments[3]:void 0;return void(0,n.iz)("visibilitychange",(function(){if(t)return void("hidden"==document.visibilityState&&e());e(document.visibilityState)}),r,i)}},1214:(e,t,r)=>{r.d(t,{em:()=>v,u5:()=>N,QU:()=>S,_L:()=>I,Gm:()=>L,Lg:()=>M,gy:()=>U,BV:()=>Q,Kf:()=>ee});var n=r(2177);const i="nr@original";var o=Object.prototype.hasOwnProperty,a=!1;function s(e,t){return e||(e=n.ee),r.inPlace=function(e,t,n,i,o){n||(n="");var a,s,c,u="-"===n.charAt(0);for(c=0;c 2?n-2:0),o=2;o {r(A[T],e,w),r(E[T],e,w)})),r(l._A,"fetch",y),t.on(y+"end",(function(e,r){var n=this;if(r){var i=r.headers.get("content-length");null!==i&&(n.rxSize=i),t.emit(y+"done",[null,r],n)}else t.emit(y+"done",[e],n)})),t}const O={},j=["pushState","replaceState"];function S(e){const t=function(e){return(e||n.ee).get("history")}(e);return!l.il||O[t.debugId]++||(O[t.debugId]=1,s(t).inPlace(window.history,j,"-")),t}var P=r(3239);const C={},R=["appendChild","insertBefore","replaceChild"];function I(e){const t=function(e){return(e||n.ee).get("jsonp")}(e);if(!l.il||C[t.debugId])return t;C[t.debugId]=!0;var r=s(t),i=/[?&](?:callback|cb)=([^&#]+)/,o=/(.*)\.([^.]+)/,a=/^(\w+)(\.|$)(.*)$/;function c(e,t){var r=e.match(a),n=r[1],i=r[3];return i?c(i,t[n]):t[n]}return r.inPlace(Node.prototype,R,"dom-"),t.on("dom-start",(function(e){!function(e){if(!e||"string"!=typeof e.nodeName||"script"!==e.nodeName.toLowerCase())return;if("function"!=typeof e.addEventListener)return;var n=(a=e.src,s=a.match(i),s?s[1]:null);var a,s;if(!n)return;var u=function(e){var t=e.match(o);if(t&&t.length>=3)return{key:t[2],parent:c(t[1],window)};return{key:e,parent:window}}(n);if("function"!=typeof u.parent[u.key])return;var d={};function f(){t.emit("jsonp-end",[],d),e.removeEventListener("load",f,(0,P.m$)(!1)),e.removeEventListener("error",l,(0,P.m$)(!1))}function l(){t.emit("jsonp-error",[],d),t.emit("jsonp-end",[],d),e.removeEventListener("load",f,(0,P.m$)(!1)),e.removeEventListener("error",l,(0,P.m$)(!1))}r.inPlace(u.parent,[u.key],"cb-",d),e.addEventListener("load",f,(0,P.m$)(!1)),e.addEventListener("error",l,(0,P.m$)(!1)),t.emit("new-jsonp",[e.src],d)}(e[0])})),t}var k=r(5763);const H={};function L(e){const t=function(e){return(e||n.ee).get("mutation")}(e);if(!l.il||H[t.debugId])return t;H[t.debugId]=!0;var r=s(t),i=k.Yu.MO;return i&&(window.MutationObserver=function(e){return this instanceof i?new i(r(e,"fn-")):i.apply(this,arguments)},MutationObserver.prototype=i.prototype),t}const z={};function M(e){const t=function(e){return(e||n.ee).get("promise")}(e);if(z[t.debugId])return t;z[t.debugId]=!0;var r=n.c,o=s(t),a=k.Yu.PR;return a&&function(){function e(r){var n=t.context(),i=o(r,"executor-",n,null,!1);const s=Reflect.construct(a,[i],e);return t.context(s).getCtx=function(){return n},s}l._A.Promise=e,Object.defineProperty(e,"name",{value:"Promise"}),e.toString=function(){return a.toString()},Object.setPrototypeOf(e,a),["all","race"].forEach((function(r){const n=a[r];e[r]=function(e){let i=!1;[...e||[]].forEach((e=>{this.resolve(e).then(a("all"===r),a(!1))}));const o=n.apply(this,arguments);return o;function a(e){return function(){t.emit("propagate",[null,!i],o,!1,!1),i=i||!e}}}})),["resolve","reject"].forEach((function(r){const n=a[r];e[r]=function(e){const r=n.apply(this,arguments);return e!==r&&t.emit("propagate",[e,!0],r,!1,!1),r}})),e.prototype=a.prototype;const n=a.prototype.then;a.prototype.then=function(){var e=this,i=r(e);i.promise=e;for(var a=arguments.length,s=new Array(a),c=0;c e())),t};function m(e,t){i.inPlace(t,["onreadystatechange"],"fn-",E)}function b(){var e=this,t=r.context(e);e.readyState>3&&!t.resolved&&(t.resolved=!0,r.emit("xhr-resolved",[],e)),i.inPlace(e,f,"fn-",E)}if(function(e,t){for(var r in e)t[r]=e[r]}(o,p),p.prototype=o.prototype,i.inPlace(p.prototype,J,"-xhr-",E),r.on("send-xhr-start",(function(e,t){m(e,t),function(e){h.push(e),a&&(y?y.then(A):u?u(A):(w=-w,x.data=w))}(t)})),r.on("open-xhr-start",m),a){var y=c&&c.resolve();if(!u&&!c){var w=1,x=document.createTextNode(w);new a(A).observe(x,{characterData:!0})}}else t.on("fn-end",(function(e){e[0]&&e[0].type===d||A()}));function A(){for(var e=0;e {r.d(t,{t:()=>n});const n=r(3325).D.ajax},6660:(e,t,r)=>{r.d(t,{A:()=>i,t:()=>n});const n=r(3325).D.jserrors,i="nr@seenError"},3081:(e,t,r)=>{r.d(t,{gF:()=>o,mY:()=>i,t9:()=>n,vz:()=>s,xS:()=>a});const n=r(3325).D.metrics,i="sm",o="cm",a="storeSupportabilityMetrics",s="storeEventMetrics"},4649:(e,t,r)=>{r.d(t,{t:()=>n});const n=r(3325).D.pageAction},7633:(e,t,r)=>{r.d(t,{Dz:()=>i,OJ:()=>a,qw:()=>o,t9:()=>n});const n=r(3325).D.pageViewEvent,i="firstbyte",o="domcontent",a="windowload"},9251:(e,t,r)=>{r.d(t,{t:()=>n});const n=r(3325).D.pageViewTiming},3614:(e,t,r)=>{r.d(t,{BST_RESOURCE:()=>i,END:()=>s,FEATURE_NAME:()=>n,FN_END:()=>u,FN_START:()=>c,PUSH_STATE:()=>d,RESOURCE:()=>o,START:()=>a});const n=r(3325).D.sessionTrace,i="bstResource",o="resource",a="-start",s="-end",c="fn"+a,u="fn"+s,d="pushState"},7836:(e,t,r)=>{r.d(t,{BODY:()=>A,CB_END:()=>E,CB_START:()=>u,END:()=>x,FEATURE_NAME:()=>i,FETCH:()=>_,FETCH_BODY:()=>v,FETCH_DONE:()=>m,FETCH_START:()=>p,FN_END:()=>c,FN_START:()=>s,INTERACTION:()=>l,INTERACTION_API:()=>d,INTERACTION_EVENTS:()=>o,JSONP_END:()=>b,JSONP_NODE:()=>g,JS_TIME:()=>T,MAX_TIMER_BUDGET:()=>a,REMAINING:()=>f,SPA_NODE:()=>h,START:()=>w,originalSetTimeout:()=>y});var n=r(5763);const i=r(3325).D.spa,o=["click","submit","keypress","keydown","keyup","change"],a=999,s="fn-start",c="fn-end",u="cb-start",d="api-ixn-",f="remaining",l="interaction",h="spaNode",g="jsonpNode",p="fetch-start",m="fetch-done",v="fetch-body-",b="jsonp-end",y=n.Yu.ST,w="-start",x="-end",A="-body",E="cb"+x,T="jsTime",_="fetch"},5938:(e,t,r)=>{r.d(t,{W:()=>o});var n=r(5763),i=r(2177);class o{constructor(e,t,r){this.agentIdentifier=e,this.aggregator=t,this.ee=i.ee.get(e,(0,n.OP)(this.agentIdentifier).isolatedBacklog),this.featureName=r,this.blocked=!1}}},9144:(e,t,r)=>{r.d(t,{j:()=>m});var n=r(3325),i=r(5763),o=r(5546),a=r(2177),s=r(7894),c=r(8e3),u=r(3960),d=r(385),f=r(50),l=r(3081),h=r(8632);function g(){const e=(0,h.gG)();["setErrorHandler","finished","addToTrace","inlineHit","addRelease","addPageAction","setCurrentRouteName","setPageViewName","setCustomAttribute","interaction","noticeError","setUserId"].forEach((t=>{e[t]=function(){for(var r=arguments.length,n=new Array(r),i=0;i 1?r-1:0),i=1;i {e.exposed&&e.api[t]&&o.push(e.api[t](...n))})),o.length>1?o:o[0]}(t,...n)}}))}var p=r(2587);function m(e){let t=arguments.length>1&&void 0!==arguments[1]?arguments[1]:{},m=arguments.length>2?arguments[2]:void 0,v=arguments.length>3?arguments[3]:void 0,{init:b,info:y,loader_config:w,runtime:x={loaderType:m},exposed:A=!0}=t;const E=(0,h.gG)();y||(b=E.init,y=E.info,w=E.loader_config),(0,i.Dg)(e,b||{}),(0,i.GE)(e,w||{}),(0,i.sU)(e,x),y.jsAttributes??={},d.v6&&(y.jsAttributes.isWorker=!0),(0,i.CX)(e,y),g();const T=function(e,t){t||(0,c.R)(e,"api");const h={};var g=a.ee.get(e),p=g.get("tracer"),m="api-",v=m+"ixn-";function b(t,r,n,o){const a=(0,i.C5)(e);return null===r?delete a.jsAttributes[t]:(0,i.CX)(e,{...a,jsAttributes:{...a.jsAttributes,[t]:r}}),x(m,n,!0,o||null===r?"session":void 0)(t,r)}function y(){}["setErrorHandler","finished","addToTrace","inlineHit","addRelease"].forEach((e=>h[e]=x(m,e,!0,"api"))),h.addPageAction=x(m,"addPageAction",!0,n.D.pageAction),h.setCurrentRouteName=x(m,"routeName",!0,n.D.spa),h.setPageViewName=function(t,r){if("string"==typeof t)return"/"!==t.charAt(0)&&(t="/"+t),(0,i.OP)(e).customTransaction=(r||"http://custom.transaction")+t,x(m,"setPageViewName",!0)()},h.setCustomAttribute=function(e,t){let r=arguments.length>2&&void 0!==arguments[2]&&arguments[2];if("string"==typeof e){if(["string","number"].includes(typeof t)||null===t)return b(e,t,"setCustomAttribute",r);(0,f.Z)("Failed to execute setCustomAttribute.\nNon-null value must be a string or number type, but a type of was provided."))}else(0,f.Z)("Failed to execute setCustomAttribute.\nName must be a string type, but a type of was provided."))},h.setUserId=function(e){if("string"==typeof e||null===e)return b("enduser.id",e,"setUserId",!0);(0,f.Z)("Failed to execute setUserId.\nNon-null value must be a string type, but a type of was provided."))},h.interaction=function(){return(new y).get()};var w=y.prototype={createTracer:function(e,t){var r={},i=this,a="function"==typeof t;return(0,o.p)(v+"tracer",[(0,s.z)(),e,r],i,n.D.spa,g),function(){if(p.emit((a?"":"no-")+"fn-start",[(0,s.z)(),i,a],r),a)try{return t.apply(this,arguments)}catch(e){throw p.emit("fn-err",[arguments,this,"string"==typeof e?new Error(e):e],r),e}finally{p.emit("fn-end",[(0,s.z)()],r)}}}};function x(e,t,r,i){return function(){return(0,o.p)(l.xS,["API/"+t+"/called"],void 0,n.D.metrics,g),i&&(0,o.p)(e+t,[(0,s.z)(),...arguments],r?null:this,i,g),r?void 0:this}}function A(){r.e(439).then(r.bind(r,7438)).then((t=>{let{setAPI:r}=t;r(e),(0,c.L)(e,"api")})).catch((()=>(0,f.Z)("Downloading runtime APIs failed...")))}return["actionText","setName","setAttribute","save","ignore","onEnd","getContext","end","get"].forEach((e=>{w[e]=x(v,e,void 0,n.D.spa)})),h.noticeError=function(e,t){"string"==typeof e&&(e=new Error(e)),(0,o.p)(l.xS,["API/noticeError/called"],void 0,n.D.metrics,g),(0,o.p)("err",[e,(0,s.z)(),!1,t],void 0,n.D.jserrors,g)},d.il?(0,u.b)((()=>A()),!0):A(),h}(e,v);return(0,h.Qy)(e,T,"api"),(0,h.Qy)(e,A,"exposed"),(0,h.EZ)("activatedFeatures",p.T),T}},3325:(e,t,r)=>{r.d(t,{D:()=>n,p:()=>i});const n={ajax:"ajax",jserrors:"jserrors",metrics:"metrics",pageAction:"page_action",pageViewEvent:"page_view_event",pageViewTiming:"page_view_timing",sessionReplay:"session_replay",sessionTrace:"session_trace",spa:"spa"},i={[n.pageViewEvent]:1,[n.pageViewTiming]:2,[n.metrics]:3,[n.jserrors]:4,[n.ajax]:5,[n.sessionTrace]:6,[n.pageAction]:7,[n.spa]:8,[n.sessionReplay]:9}}},n={};function i(e){var t=n[e];if(void 0!==t)return t.exports;var o=n[e]={exports:{}};return r[e](o,o.exports,i),o.exports}i.m=r,i.d=(e,t)=>{for(var r in t)i.o(t,r)&&!i.o(e,r)&&Object.defineProperty(e,r,{enumerable:!0,get:t[r]})},i.f={},i.e=e=>Promise.all(Object.keys(i.f).reduce(((t,r)=>(i.f[r](e,t),t)),[])),i.u=e=>(({78:"page_action-aggregate",147:"metrics-aggregate",242:"session-manager",317:"jserrors-aggregate",348:"page_view_timing-aggregate",412:"lazy-feature-loader",439:"async-api",538:"recorder",590:"session_replay-aggregate",675:"compressor",733:"session_trace-aggregate",786:"page_view_event-aggregate",873:"spa-aggregate",898:"ajax-aggregate"}[e]||e)+"."+{78:"ac76d497",147:"3dc53903",148:"1a20d5fe",242:"2a64278a",317:"49e41428",348:"bd6de33a",412:"2f55ce66",439:"30bd804e",538:"1b18459f",590:"cf0efb30",675:"ae9f91a8",733:"83105561",786:"06482edd",860:"03a8b7a5",873:"e6b09d52",898:"998ef92b"}[e]+"-1.236.0.min.js"),i.o=(e,t)=>Object.prototype.hasOwnProperty.call(e,t),e={},t="NRBA:",i.l=(r,n,o,a)=>{if(e[r])e[r].push(n);else{var s,c;if(void 0!==o)for(var u=document.getElementsByTagName("script"),d=0;d {s.onerror=s.onload=null,clearTimeout(h);var i=e[r];if(delete e[r],s.parentNode&&s.parentNode.removeChild(s),i&&i.forEach((e=>e(n))),t)return t(n)},h=setTimeout(l.bind(null,void 0,{type:"timeout",target:s}),12e4);s.onerror=l.bind(null,s.onerror),s.onload=l.bind(null,s.onload),c&&document.head.appendChild(s)}},i.r=e=>{"undefined"!=typeof Symbol&&Symbol.toStringTag&&Object.defineProperty(e,Symbol.toStringTag,{value:"Module"}),Object.defineProperty(e,"__esModule",{value:!0})},i.j=364,i.p="https://js-agent.newrelic.com/",(()=>{var e={364:0,953:0};i.f.j=(t,r)=>{var n=i.o(e,t)?e[t]:void 0;if(0!==n)if(n)r.push(n[2]);else{var o=new Promise(((r,i)=>n=e[t]=[r,i]));r.push(n[2]=o);var a=i.p+i.u(t),s=new Error;i.l(a,(r=>{if(i.o(e,t)&&(0!==(n=e[t])&&(e[t]=void 0),n)){var o=r&&("load"===r.type?"missing":r.type),a=r&&r.target&&r.target.src;s.message="Loading chunk "+t+" failed.\n("+o+": "+a+")",s.name="ChunkLoadError",s.type=o,s.request=a,n[1](s)}}),"chunk-"+t,t)}};var t=(t,r)=>{var n,o,[a,s,c]=r,u=0;if(a.some((t=>0!==e[t]))){for(n in s)i.o(s,n)&&(i.m[n]=s[n]);if(c)c(i)}for(t&&t(r);u {i.r(o);var e=i(3325),t=i(5763);const r=Object.values(e.D);function n(e){const n={};return r.forEach((r=>{n[r]=function(e,r){return!1!==(0,t.Mt)(r,"".concat(e,".enabled"))}(r,e)})),n}var a=i(9144);var s=i(5546),c=i(385),u=i(8e3),d=i(5938),f=i(3960),l=i(50);class h extends d.W{constructor(e,t,r){let n=!(arguments.length>3&&void 0!==arguments[3])||arguments[3];super(e,t,r),this.auto=n,this.abortHandler,this.featAggregate,this.onAggregateImported,n&&(0,u.R)(e,r)}importAggregator(){let e=arguments.length>0&&void 0!==arguments[0]?arguments[0]:{};if(this.featAggregate||!this.auto)return;const r=c.il&&!0===(0,t.Mt)(this.agentIdentifier,"privacy.cookies_enabled");let n;this.onAggregateImported=new Promise((e=>{n=e}));const o=async()=>{let t;try{if(r){const{setupAgentSession:e}=await Promise.all([i.e(860),i.e(242)]).then(i.bind(i,3228));t=e(this.agentIdentifier)}}catch(e){(0,l.Z)("A problem occurred when starting up session manager. This page will not start or extend any session.",e)}try{if(!this.shouldImportAgg(this.featureName,t))return void(0,u.L)(this.agentIdentifier,this.featureName);const{lazyFeatureLoader:r}=await i.e(412).then(i.bind(i,8582)),{Aggregate:o}=await r(this.featureName,"aggregate");this.featAggregate=new o(this.agentIdentifier,this.aggregator,e),n(!0)}catch(e){(0,l.Z)("Downloading and initializing ".concat(this.featureName," failed..."),e),this.abortHandler?.(),n(!1)}};c.il?(0,f.b)((()=>o()),!0):o()}shouldImportAgg(r,n){return r!==e.D.sessionReplay||!1!==(0,t.Mt)(this.agentIdentifier,"session_trace.enabled")&&(!!n?.isNew||!!n?.state.sessionReplay)}}var g=i(7633),p=i(7894);class m extends h{static featureName=g.t9;constructor(r,n){let i=!(arguments.length>2&&void 0!==arguments[2])||arguments[2];if(super(r,n,g.t9,i),("undefined"==typeof PerformanceNavigationTiming||c.Tt)&&"undefined"!=typeof PerformanceTiming){const n=(0,t.OP)(r);n[g.Dz]=Math.max(Date.now()-n.offset,0),(0,f.K)((()=>n[g.qw]=Math.max((0,p.z)()-n[g.Dz],0))),(0,f.b)((()=>{const t=(0,p.z)();n[g.OJ]=Math.max(t-n[g.Dz],0),(0,s.p)("timing",["load",t],void 0,e.D.pageViewTiming,this.ee)}))}this.importAggregator()}}var v=i(1117),b=i(1284);class y extends v.w{constructor(e){super(e),this.aggregatedData={}}store(e,t,r,n,i){var o=this.getBucket(e,t,r,i);return o.metrics=function(e,t){t||(t={count:0});return t.count+=1,(0,b.D)(e,(function(e,r){t[e]=w(r,t[e])})),t}(n,o.metrics),o}merge(e,t,r,n,i){var o=this.getBucket(e,t,n,i);if(o.metrics){var a=o.metrics;a.count+=r.count,(0,b.D)(r,(function(e,t){if("count"!==e){var n=a[e],i=r[e];i&&!i.c?a[e]=w(i.t,n):a[e]=function(e,t){if(!t)return e;t.c||(t=x(t.t));return t.min=Math.min(e.min,t.min),t.max=Math.max(e.max,t.max),t.t+=e.t,t.sos+=e.sos,t.c+=e.c,t}(i,a[e])}}))}else o.metrics=r}storeMetric(e,t,r,n){var i=this.getBucket(e,t,r);return i.stats=w(n,i.stats),i}getBucket(e,t,r,n){this.aggregatedData[e]||(this.aggregatedData[e]={});var i=this.aggregatedData[e][t];return i||(i=this.aggregatedData[e][t]={params:r||{}},n&&(i.custom=n)),i}get(e,t){return t?this.aggregatedData[e]&&this.aggregatedData[e][t]:this.aggregatedData[e]}take(e){for(var t={},r="",n=!1,i=0;i t.max&&(t.max=e),e 2&&void 0!==arguments[2])||arguments[2];super(e,r,j.t,n),c.il&&((0,t.OP)(e).initHidden=Boolean("hidden"===document.visibilityState),(0,N.N)((()=>(0,s.p)("docHidden",[(0,p.z)()],void 0,j.t,this.ee)),!0),(0,O.bP)("pagehide",(()=>(0,s.p)("winPagehide",[(0,p.z)()],void 0,j.t,this.ee))),this.importAggregator())}}var P=i(3081);class C extends h{static featureName=P.t9;constructor(e,t){let r=!(arguments.length>2&&void 0!==arguments[2])||arguments[2];super(e,t,P.t9,r),this.importAggregator()}}var R,I=i(2210),k=i(1214),H=i(2177),L={};try{R=localStorage.getItem("__nr_flags").split(","),console&&"function"==typeof console.log&&(L.console=!0,-1!==R.indexOf("dev")&&(L.dev=!0),-1!==R.indexOf("nr_dev")&&(L.nrDev=!0))}catch(e){}function z(e){try{L.console&&z(e)}catch(e){}}L.nrDev&&H.ee.on("internal-error",(function(e){z(e.stack)})),L.dev&&H.ee.on("fn-err",(function(e,t,r){z(r.stack)})),L.dev&&(z("NR AGENT IN DEVELOPMENT MODE"),z("flags: "+(0,b.D)(L,(function(e,t){return e})).join(", ")));var M=i(6660);class B extends h{static featureName=M.t;constructor(r,n){let i=!(arguments.length>2&&void 0!==arguments[2])||arguments[2];super(r,n,M.t,i),this.skipNext=0;try{this.removeOnAbort=new AbortController}catch(e){}const o=this;o.ee.on("fn-start",(function(e,t,r){o.abortHandler&&(o.skipNext+=1)})),o.ee.on("fn-err",(function(t,r,n){o.abortHandler&&!n[M.A]&&((0,I.X)(n,M.A,(function(){return!0})),this.thrown=!0,(0,s.p)("err",[n,(0,p.z)()],void 0,e.D.jserrors,o.ee))})),o.ee.on("fn-end",(function(){o.abortHandler&&!this.thrown&&o.skipNext>0&&(o.skipNext-=1)})),o.ee.on("internal-error",(function(t){(0,s.p)("ierr",[t,(0,p.z)(),!0],void 0,e.D.jserrors,o.ee)})),this.origOnerror=c._A.onerror,c._A.onerror=this.onerrorHandler.bind(this),c._A.addEventListener("unhandledrejection",(t=>{const r=function(e){let t="Unhandled Promise Rejection: ";if(e instanceof Error)try{return e.message=t+e.message,e}catch(t){return e}if(void 0===e)return new Error(t);try{return new Error(t+(0,D.P)(e))}catch(e){return new Error(t)}}(t.reason);(0,s.p)("err",[r,(0,p.z)(),!1,{unhandledPromiseRejection:1}],void 0,e.D.jserrors,this.ee)}),(0,O.m$)(!1,this.removeOnAbort?.signal)),(0,k.gy)(this.ee),(0,k.BV)(this.ee),(0,k.em)(this.ee),(0,t.OP)(r).xhrWrappable&&(0,k.Kf)(this.ee),this.abortHandler=this.#e,this.importAggregator()}#e(){this.removeOnAbort?.abort(),this.abortHandler=void 0}onerrorHandler(t,r,n,i,o){"function"==typeof this.origOnerror&&this.origOnerror(...arguments);try{this.skipNext?this.skipNext-=1:(0,s.p)("err",[o||new F(t,r,n),(0,p.z)()],void 0,e.D.jserrors,this.ee)}catch(t){try{(0,s.p)("ierr",[t,(0,p.z)(),!0],void 0,e.D.jserrors,this.ee)}catch(e){}}return!1}}function F(e,t,r){this.message=e||"Uncaught error with no additional information",this.sourceURL=t,this.line=r}let U=1;const q="nr@id";function G(e){const t=typeof e;return!e||"object"!==t&&"function"!==t?-1:e===c._A?0:(0,I.X)(e,q,(function(){return U++}))}function V(e){if("string"==typeof e&&e.length)return e.length;if("object"==typeof e){if("undefined"!=typeof ArrayBuffer&&e instanceof ArrayBuffer&&e.byteLength)return e.byteLength;if("undefined"!=typeof Blob&&e instanceof Blob&&e.size)return e.size;if(!("undefined"!=typeof FormData&&e instanceof FormData))try{return(0,D.P)(e).length}catch(e){return}}}var X=i(7243);class W{constructor(e){this.agentIdentifier=e,this.generateTracePayload=this.generateTracePayload.bind(this),this.shouldGenerateTrace=this.shouldGenerateTrace.bind(this)}generateTracePayload(e){if(!this.shouldGenerateTrace(e))return null;var r=(0,t.DL)(this.agentIdentifier);if(!r)return null;var n=(r.accountID||"").toString()||null,i=(r.agentID||"").toString()||null,o=(r.trustKey||"").toString()||null;if(!n||!i)return null;var a=(0,_.M)(),s=(0,_.Ht)(),c=Date.now(),u={spanId:a,traceId:s,timestamp:c};return(e.sameOrigin||this.isAllowedOrigin(e)&&this.useTraceContextHeadersForCors())&&(u.traceContextParentHeader=this.generateTraceContextParentHeader(a,s),u.traceContextStateHeader=this.generateTraceContextStateHeader(a,c,n,i,o)),(e.sameOrigin&&!this.excludeNewrelicHeader()||!e.sameOrigin&&this.isAllowedOrigin(e)&&this.useNewrelicHeaderForCors())&&(u.newrelicHeader=this.generateTraceHeader(a,s,c,n,i,o)),u}generateTraceContextParentHeader(e,t){return"00-"+t+"-"+e+"-01"}generateTraceContextStateHeader(e,t,r,n,i){return i+"@nr=0-1-"+r+"-"+n+"-"+e+"----"+t}generateTraceHeader(e,t,r,n,i,o){if(!("function"==typeof c._A?.btoa))return null;var a={v:[0,1],d:{ty:"Browser",ac:n,ap:i,id:e,tr:t,ti:r}};return o&&n!==o&&(a.d.tk=o),btoa((0,D.P)(a))}shouldGenerateTrace(e){return this.isDtEnabled()&&this.isAllowedOrigin(e)}isAllowedOrigin(e){var r=!1,n={};if((0,t.Mt)(this.agentIdentifier,"distributed_tracing")&&(n=(0,t.P_)(this.agentIdentifier).distributed_tracing),e.sameOrigin)r=!0;else if(n.allowed_origins instanceof Array)for(var i=0;i 2&&void 0!==arguments[2])||arguments[2];super(r,n,Z.t,i),(0,t.OP)(r).xhrWrappable&&(this.dt=new W(r),this.handler=(e,t,r,n)=>(0,s.p)(e,t,r,n,this.ee),(0,k.u5)(this.ee),(0,k.Kf)(this.ee),function(r,n,i,o){function a(e){var t=this;t.totalCbs=0,t.called=0,t.cbTime=0,t.end=E,t.ended=!1,t.xhrGuids={},t.lastSize=null,t.loadCaptureCalled=!1,t.params=this.params||{},t.metrics=this.metrics||{},e.addEventListener("load",(function(r){_(t,e)}),(0,O.m$)(!1)),c.IF||e.addEventListener("progress",(function(e){t.lastSize=e.loaded}),(0,O.m$)(!1))}function s(e){this.params={method:e[0]},T(this,e[1]),this.metrics={}}function u(e,n){var i=(0,t.DL)(r);i.xpid&&this.sameOrigin&&n.setRequestHeader("X-NewRelic-ID",i.xpid);var a=o.generateTracePayload(this.parsedOrigin);if(a){var s=!1;a.newrelicHeader&&(n.setRequestHeader("newrelic",a.newrelicHeader),s=!0),a.traceContextParentHeader&&(n.setRequestHeader("traceparent",a.traceContextParentHeader),a.traceContextStateHeader&&n.setRequestHeader("tracestate",a.traceContextStateHeader),s=!0),s&&(this.dt=a)}}function d(e,t){var r=this.metrics,i=e[0],o=this;if(r&&i){var a=V(i);a&&(r.txSize=a)}this.startTime=(0,p.z)(),this.listener=function(e){try{"abort"!==e.type||o.loadCaptureCalled||(o.params.aborted=!0),("load"!==e.type||o.called===o.totalCbs&&(o.onloadCalled||"function"!=typeof t.onload)&&"function"==typeof o.end)&&o.end(t)}catch(e){try{n.emit("internal-error",[e])}catch(e){}}};for(var s=0;s 1?e[1]=i:e.push(i)}else e[0]&&e[0].headers&&s(e[0].headers,n)&&(this.dt=n);function s(e,t){var r=!1;return t.newrelicHeader&&(e.set("newrelic",t.newrelicHeader),r=!0),t.traceContextParentHeader&&(e.set("traceparent",t.traceContextParentHeader),t.traceContextStateHeader&&e.set("tracestate",t.traceContextStateHeader),r=!0),r}}function x(e,t){this.params={},this.metrics={},this.startTime=(0,p.z)(),this.dt=t,e.length>=1&&(this.target=e[0]),e.length>=2&&(this.opts=e[1]);var r,n=this.opts||{},i=this.target;"string"==typeof i?r=i:"object"==typeof i&&i instanceof Y?r=i.url:c._A?.URL&&"object"==typeof i&&i instanceof URL&&(r=i.href),T(this,r);var o=(""+(i&&i instanceof Y&&i.method||n.method||"GET")).toUpperCase();this.params.method=o,this.txSize=V(n.body)||0}function A(t,r){var n;this.endTime=(0,p.z)(),this.params||(this.params={}),this.params.status=r?r.status:0,"string"==typeof this.rxSize&&this.rxSize.length>0&&(n=+this.rxSize);var o={txSize:this.txSize,rxSize:n,duration:(0,p.z)()-this.startTime};i("xhr",[this.params,o,this.startTime,this.endTime,"fetch"],this,e.D.ajax)}function E(t){var r=this.params,n=this.metrics;if(!this.ended){this.ended=!0;for(var o=0;o 2&&void 0!==arguments[2])||arguments[2];super(e,t,we.t,r),this.importAggregator()}}new class{constructor(e){let t=arguments.length>1&&void 0!==arguments[1]?arguments[1]:(0,_.ky)(16);c._A?(this.agentIdentifier=t,this.sharedAggregator=new y({agentIdentifier:this.agentIdentifier}),this.features={},this.desiredFeatures=new Set(e.features||[]),this.desiredFeatures.add(m),Object.assign(this,(0,a.j)(this.agentIdentifier,e,e.loaderType||"agent")),this.start()):(0,l.Z)("Failed to initial the agent. Could not determine the runtime environment.")}get config(){return{info:(0,t.C5)(this.agentIdentifier),init:(0,t.P_)(this.agentIdentifier),loader_config:(0,t.DL)(this.agentIdentifier),runtime:(0,t.OP)(this.agentIdentifier)}}start(){const t="features";try{const r=n(this.agentIdentifier),i=[...this.desiredFeatures];i.sort(((t,r)=>e.p[t.featureName]-e.p[r.featureName])),i.forEach((t=>{if(r[t.featureName]||t.featureName===e.D.pageViewEvent){const n=function(t){switch(t){case e.D.ajax:return[e.D.jserrors];case e.D.sessionTrace:return[e.D.ajax,e.D.pageViewEvent];case e.D.sessionReplay:return[e.D.sessionTrace];case e.D.pageViewTiming:return[e.D.pageViewEvent];default:return[]}}(t.featureName);n.every((e=>r[e]))||(0,l.Z)("".concat(t.featureName," is enabled but one or more dependent features has been disabled (").concat((0,D.P)(n),"). This may cause unintended consequences or missing data...")),this.features[t.featureName]=new t(this.agentIdentifier,this.sharedAggregator)}})),(0,T.Qy)(this.agentIdentifier,this.features,t)}catch(e){(0,l.Z)("Failed to initialize all enabled instrument classes (agent aborted) -",e);for(const e in this.features)this.features[e].abortHandler?.();const r=(0,T.fP)();return delete r.initializedAgents[this.agentIdentifier]?.api,delete r.initializedAgents[this.agentIdentifier]?.[t],delete this.sharedAggregator,r.ee?.abort(),delete r.ee?.get(this.agentIdentifier),!1}}}({features:[J,m,S,class extends h{static featureName=oe;constructor(t,r){if(super(t,r,oe,!(arguments.length>2&&void 0!==arguments[2])||arguments[2]),!c.il)return;const n=this.ee;let i;(0,k.QU)(n),this.eventsEE=(0,k.em)(n),this.eventsEE.on(se,(function(e,t){this.bstStart=(0,p.z)()})),this.eventsEE.on(ae,(function(t,r){(0,s.p)("bst",[t[0],r,this.bstStart,(0,p.z)()],void 0,e.D.sessionTrace,n)})),n.on(ce+ne,(function(e){this.time=(0,p.z)(),this.startPath=location.pathname+location.hash})),n.on(ce+ie,(function(t){(0,s.p)("bstHist",[location.pathname+location.hash,this.startPath,this.time],void 0,e.D.sessionTrace,n)}));try{i=new PerformanceObserver((t=>{const r=t.getEntries();(0,s.p)(te,[r],void 0,e.D.sessionTrace,n)})),i.observe({type:re,buffered:!0})}catch(e){}this.importAggregator({resourceObserver:i})}},C,xe,B,class extends h{static featureName=de;constructor(e,r){if(super(e,r,de,!(arguments.length>2&&void 0!==arguments[2])||arguments[2]),!c.il)return;if(!(0,t.OP)(e).xhrWrappable)return;try{this.removeOnAbort=new AbortController}catch(e){}let n,i=0;const o=this.ee.get("tracer"),a=(0,k._L)(this.ee),s=(0,k.Lg)(this.ee),u=(0,k.BV)(this.ee),d=(0,k.Kf)(this.ee),f=this.ee.get("events"),l=(0,k.u5)(this.ee),h=(0,k.QU)(this.ee),g=(0,k.Gm)(this.ee);function m(e,t){h.emit("newURL",[""+window.location,t])}function v(){i++,n=window.location.hash,this[ve]=(0,p.z)()}function b(){i--,window.location.hash!==n&&m(0,!0);var e=(0,p.z)();this[pe]=~~this[pe]+e-this[ve],this[ye]=e}function y(e,t){e.on(t,(function(){this[t]=(0,p.z)()}))}this.ee.on(ve,v),s.on(be,v),a.on(be,v),this.ee.on(ye,b),s.on(ge,b),a.on(ge,b),this.ee.buffer([ve,ye,"xhr-resolved"],this.featureName),f.buffer([ve],this.featureName),u.buffer(["setTimeout"+le,"clearTimeout"+fe,ve],this.featureName),d.buffer([ve,"new-xhr","send-xhr"+fe],this.featureName),l.buffer([me+fe,me+"-done",me+he+fe,me+he+le],this.featureName),h.buffer(["newURL"],this.featureName),g.buffer([ve],this.featureName),s.buffer(["propagate",be,ge,"executor-err","resolve"+fe],this.featureName),o.buffer([ve,"no-"+ve],this.featureName),a.buffer(["new-jsonp","cb-start","jsonp-error","jsonp-end"],this.featureName),y(l,me+fe),y(l,me+"-done"),y(a,"new-jsonp"),y(a,"jsonp-end"),y(a,"cb-start"),h.on("pushState-end",m),h.on("replaceState-end",m),window.addEventListener("hashchange",m,(0,O.m$)(!0,this.removeOnAbort?.signal)),window.addEventListener("load",m,(0,O.m$)(!0,this.removeOnAbort?.signal)),window.addEventListener("popstate",(function(){m(0,i>1)}),(0,O.m$)(!0,this.removeOnAbort?.signal)),this.abortHandler=this.#e,this.importAggregator()}#e(){this.removeOnAbort?.abort(),this.abortHandler=void 0}}],loaderType:"spa"})})(),window.NRBA=o})(); window.jQuery || document.write(' ') CKEDITOR_BASEPATH='https://f1000research.com/js/vendor/ckeditor/' window.reactTheme = 'research'; window.MathJax = { CommonHTML: { linebreaks: { automatic: true } }, 'HTML-CSS': { linebreaks: { automatic: true } }, SVG: { linebreaks: { automatic: true } }, AuthorInit: function() { MathJax.Hub.Register.MessageHook('End Process', function () { let timeout = false; // holder for timeout id const delay = 250; // delay after event is "complete" to run callback const reflowMath = function() { const dispFormulas = document.querySelectorAll('.disp-formula.panel'); if (!dispFormulas) { return; } for (const dispFormula of dispFormulas) { const child = dispFormula.querySelector('.MathJax_Preview').nextSibling.firstChild; const isMultiline = MathJax.Hub.getAllJax(dispFormula)[0].root.isMultiline; if (dispFormula.offsetWidth < child.offsetWidth || isMultiline) { MathJax.Hub.Queue(['Rerender', MathJax.Hub, dispFormula]); } } }; window.addEventListener('resize', function() { clearTimeout(timeout); // clear the timeout timeout = setTimeout(reflowMath, delay); // start timing for event "completion" }); }); }, }; if (window.location.hash == '#_=_'){ window.location = window.location.href.split('#')[0] } !function(f,b,e,v,n,t,s){if(f.fbq)return;n=f.fbq=function() {n.callMethod? n.callMethod.apply(n,arguments):n.queue.push(arguments)} ;if(!f._fbq)f._fbq=n; n.push=n;n.loaded=!0;n.version='2.0';n.queue=[];t=b.createElement(e);t.async=!0; t.src=v;s=b.getElementsByTagName(e)[0];s.parentNode.insertBefore(t,s)}(window, document,'script','https://connect.facebook.net/en_US/fbevents.js'); fbq('init', '1641728616063202'); fbq('track', "PixelInitialized", {}); (function(h,o,t,j,a,r){ h.hj=h.hj||function(){(h.hj.q=h.hj.q||[]).push(arguments)}; h._hjSettings={hjid:2318163,hjsv:6}; a=o.getElementsByTagName('head')[0]; r=o.createElement('script');r.async=1; r.src=t+h._hjSettings.hjid+j+h._hjSettings.hjsv; a.appendChild(r); })(window,document,'https://static.hotjar.com/c/hotjar-','.js?sv='); search file_upload Submit your research search menu close search Browse Gateways & Collections How to Publish Submit your Research My Submissions Article Guidelines Article Guidelines (New Versions) Open Data, Software and Code Guidelines Open Data and Accessible Source Materials Guidelines (HSS) Open Data, Software and Code Guidelines (PSE) Prepublication Checks Production Process Posters and Slides Guidelines Document Guidelines Article Processing Charges Peer Review Finding Article Reviewers About How it Works For Reviewers Our Advisors Policies Glossary FAQs For Developers Newsroom Contact My Research Submissions Content and Tracking Alerts My Details Sign In file_upload Submit your research { "@context": "https://schema.org", "@type": "ScholarlyArticle", "mainEntityOfPage": { "@type": "WebPage", "@id": "https://f1000research.com/articles/12-945" }, "headline": "Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric", "datePublished": "2023-08-07T17:24:11", "dateModified": "2024-07-10T12:33:46", "author": [ { "@type": "Person", "name": "James Fleming" }, { "@type": "Person", "name": "Pia Merete Eriksen" }, { "@type": "Person", "name": "Torsten Hugo Struck" } ], "publisher": { "@type": "Organization", "name": "F1000Research", "logo": { "@type": "ImageObject", "url": "https://f1000research.com/img/AMP/F1000Research_image.png", "height": 480, "width": 60 } }, "image": { "@type": "ImageObject", "url": "https://f1000research.com/img/AMP/F1000Research_image.png", "height": 1200, "width": 150 }, "description": " Background: The phylogenetic bootstrap, first proposed by Felsenstein in 1985, is a critically important statistical method in assessing the robusticity of phylogenetic datasets. Core to its concept was the use of pseudo sampling - assessing the data by generating new replicates derived from the initial dataset that was used to generate the phylogeny. In this way, phylogenetic support metrics could overcome the lack of perfect, infinite data. With infinite data, however, it is possible to sample smaller replicates directly from the data to obtain both the phylogeny and its statistical robusticity in the same analysis. Due to the growth of whole genome sequencing, the depth and breadth of our datasets have greatly expanded and are set to only expand further. With genome-scale datasets comprising thousands of genes, we can now obtain a proxy for infinite data. Accordingly, we can potentially abandon the notion of pseudo sampling and instead randomly sample small subsets of genes from the thousands of genes in our analyses. Methods: We introduce Scoutknife, a jackknife-style subsampling implementation that generates 100 datasets by randomly sampling a small number of genes from an initial large-gene dataset to jointly establish both a phylogenetic hypothesis and assess its robusticity. We assess its effectiveness by using 18 previously published datasets and 100 simulation studies. Results: We show that Scoutknife is conservative and informative as to conflicts and incongruence across the whole genome, without the need for subsampling based on traditional model selection criteria. Conclusions: Scoutknife reliably achieves comparable results to selecting the best genes on both real and simulation datasets, while being resistant to the potential biases caused by selecting for model fit. As the amount of genome data grows, it becomes an even more exciting option to assess the robusticity of phylogenetic hypotheses." } { "@context": "http://schema.org", "@type": "BreadcrumbList", "itemListElement": [ { "@type": "ListItem", "position": "1", "item": { "@id": "https://f1000research.com/", "name": "Home" } }, { "@type": "ListItem", "position": "2", "item": { "@id": "https://f1000research.com/browse/articles", "name": "Browse" } }, { "@type": "ListItem", "position": "3", "item": { "@id": "https://f1000research.com/articles/12-945", "name": "Scoutknife: A naïve, whole genome informed phylogenetic robusticity..." } } ] } Home Browse Scoutknife: A naïve, whole genome informed phylogenetic robusticity... ALL Metrics - Views Downloads Get PDF Get XML Cite How to cite this article Fleming J, Eriksen PM and Struck TH. Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric [version 2; peer review: 3 approved] . F1000Research 2024, 12 :945 ( https://doi.org/10.12688/f1000research.139356.2 ) NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article. Close Copy Citation Details Export Export Citation Sciwheel EndNote Ref. Manager Bibtex ProCite Sente EXPORT Select a format first Track Share ▬ ✚ Research Article Revised Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric [version 2; peer review: 3 approved] James Fleming https://orcid.org/0000-0001-9987-3984 1 , Pia Merete Eriksen https://orcid.org/0000-0003-1832-4794 1 , Torsten Hugo Struck 1 James Fleming https://orcid.org/0000-0001-9987-3984 1 , Pia Merete Eriksen https://orcid.org/0000-0003-1832-4794 1 , Torsten Hugo Struck 1 PUBLISHED 10 Jul 2024 Author details Author details 1 Natural History Museum, Universitetet i Oslo, Oslo, Oslo, 0562, Norway James Fleming Roles: Conceptualization, Formal Analysis, Investigation, Methodology, Software, Writing – Original Draft Preparation Pia Merete Eriksen Roles: Formal Analysis, Investigation, Writing – Review & Editing Torsten Hugo Struck Roles: Conceptualization, Funding Acquisition, Methodology, Resources, Supervision, Writing – Review & Editing OPEN PEER REVIEW DETAILS REVIEWER STATUS This article is included in the Bioinformatics gateway. This article is included in the Evolutionary Bioinformatics collection. Abstract Background: The phylogenetic bootstrap, first proposed by Felsenstein in 1985, is a critically important statistical method in assessing the robusticity of phylogenetic datasets. Core to its concept was the use of pseudo sampling - assessing the data by generating new replicates derived from the initial dataset that was used to generate the phylogeny. In this way, phylogenetic support metrics could overcome the lack of perfect, infinite data. With infinite data, however, it is possible to sample smaller replicates directly from the data to obtain both the phylogeny and its statistical robusticity in the same analysis. Due to the growth of whole genome sequencing, the depth and breadth of our datasets have greatly expanded and are set to only expand further. With genome-scale datasets comprising thousands of genes, we can now obtain a proxy for infinite data. Accordingly, we can potentially abandon the notion of pseudo sampling and instead randomly sample small subsets of genes from the thousands of genes in our analyses. Methods: We introduce Scoutknife, a jackknife-style subsampling implementation that generates 100 datasets by randomly sampling a small number of genes from an initial large-gene dataset to jointly establish both a phylogenetic hypothesis and assess its robusticity. We assess its effectiveness by using 18 previously published datasets and 100 simulation studies. Results: We show that Scoutknife is conservative and informative as to conflicts and incongruence across the whole genome, without the need for subsampling based on traditional model selection criteria. Conclusions: Scoutknife reliably achieves comparable results to selecting the best genes on both real and simulation datasets, while being resistant to the potential biases caused by selecting for model fit. As the amount of genome data grows, it becomes an even more exciting option to assess the robusticity of phylogenetic hypotheses. READ ALL READ LESS Keywords phylogenetics, bootstrapping, software Corresponding Author(s) James Fleming ( [email protected] ) Close Corresponding author: James Fleming Competing interests: No competing interests were disclosed. Grant information: This work was funded by the Research Council Norway (project number 300587 to T.H.S.). T.H.S. received additional support by the Norwegian Metacenter for Computational Science (NOTUR; project numbers NN9408K, and NS9408K). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Copyright: © 2024 Fleming J et al . This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. How to cite: Fleming J, Eriksen PM and Struck TH. Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric [version 2; peer review: 3 approved] . F1000Research 2024, 12 :945 ( https://doi.org/10.12688/f1000research.139356.2 ) First published: 07 Aug 2023, 12 :945 ( https://doi.org/10.12688/f1000research.139356.1 ) Latest published: 10 Jul 2024, 12 :945 ( https://doi.org/10.12688/f1000research.139356.2 ) Revised Amendments from Version 1 This revision presents a few key changes to the manuscript: - The introduction is significantly expanded, including a clearer discussion of the Scoutknife sampling process alongside discussion of previous implementations of gene-wise bootstrapping and subsampling, to illustrate the ways in which Scoutknife is similar and distinct from this prior work. - In response to the reviewer's comments, the Github containing the Scoutknife package is also greatly expanded - the Scoutknife package now includes an 'auto' function, which calculates the ideal sample size for an input directory of fasta files alongside a consensus tree maker. This revision presents a few key changes to the manuscript: - The introduction is significantly expanded, including a clearer discussion of the Scoutknife sampling process alongside discussion of previous implementations of gene-wise bootstrapping and subsampling, to illustrate the ways in which Scoutknife is similar and distinct from this prior work. - In response to the reviewer's comments, the Github containing the Scoutknife package is also greatly expanded - the Scoutknife package now includes an 'auto' function, which calculates the ideal sample size for an input directory of fasta files alongside a consensus tree maker. See the authors' detailed response to the review by Paul Zaharias See the authors' detailed response to the review by Xianzhao Kan READ REVIEWER RESPONSES Introduction The genomics revolution completely altered our understanding of phylogeny - the study of the relationships between organismal groups. By combining molecular and morphological data, our picture of the evolution of life has become clearer than ever before. 1 , 2 We are now in the process of entering the next phase of the genomics revolution, however, where beyond single or multi-gene datasets, researchers are now able to accurately sequence whole genomes from multiple species with relative ease. 3 – 5 This new era of “big data”, properly leveraged, promises to revolutionise our understanding of phylogenetics in the same way our prior understanding was revolutionised by the discovery of genomics itself. However, appropriately handling this new data is key to unlocking its potential. First, the robustness or statistical significance of these new results must be appropriately assessed. Second, assurances must be provided that the phylogenies reflect the actual biological processes and are not being misled by reconstructive biases. 1 , 6 , 7 The robustness and reliability of a phylogenetic topology and its branches can be quantified in a number of ways, such as through Bayesian posterior probabilities 8 or the Likelihood Ratio Test family of support values. 9 , 10 One of the most common, however, is the bootstrap support value. 11 A measure of statistical robustness, the bootstrap was first applied to phylogenetics by Felsenstein in 1985. In its implementation, the phylogeny is reconstructed from the limited source dataset and the bootstrap creates multiple pseudo-replicates of the source datasets – effectively multiplying the signal of some sites in the datasets and removing the signal of others ( Figure 1A ). A variation of this approach is the generation of pseudo-samples by jackknifing – resampling only a fraction of the sites (e.g., 60% or 80%) from the source dataset. 12 This measures robustness, assessing how many of the sites in the source dataset support the final phylogeny – or more specifically its branches – and thereby whether there is a broader consensus for the proposed most likely topology – or for a particular branch – amongst the source dataset’s component sites. 13 These pseudo-replicates can be thought of as a measure of the evenness of signal across a phylogenetic dataset. In each pseudo-sample, certain sites are removed, while other sites remain (jackknifing) or are duplicated (bootstrapping), which has the effect of muting the removed sites whilst amplifying the signal of the other sites. Bootstrapping is particularly useful where data is limited, such as in single or limited gene datasets, where pseudo-replicates can greatly proportionally increase the effective size, and thereby statistical power, of the analysis. 11 It was originally implemented as a surrogate for a robust statistical sampling procedure from theoretically unlimited data. 14 In the case of unlimited data, one could generate random samples from these data, then generate the tree of each sample and determine the overall phylogeny by including measurements of the robustness across all generated trees ( Figure 1B ). Figure 1. A figure showing a bootstrap pseudosampling process (Panel A) and a Scoutknife sampling process (Panel C), with the theoretical unlimited data jackknife sample in the middle (Panel B). Note that Scoutknife bears more similarity to unlimited data sampling than a traditional bootstrap. Scoutknife may not take the same gene twice within the same sample but may take the same gene multiple times between samples – see Scoutknife replicate #1 and #2, which both sample gene #97. The structure of this figure is based upon Hillis et al. (1996), Chapter 11, page 508, Figure 33. 18 In the modern era, genome-scale data is being generated for a rapidly increasing number of species across the tree of life. 4 , 15 – 17 When data is plentiful, the reliance on pseudo-samples becomes less necessary. With thousands of genes and millions to billions of base pairs on the horizon for phylogenetic analyses, one can safely assume that the theoretical assumption of unlimited data is not violated. Accordingly, the data can be repeatedly sampled directly, trees reconstructed and the phylogeny and statistical support determined ( Figure 1C ) as outlined directly from the aforementioned unlimited data. It is this assumption that forms the philosophical basis of the Scoutknife approach. At the same time, however, the reconstruction of the species history can be challenging due to either methodological incongruence (i.e., not all genes contain information about the species history that we can correctly decipher) or biological incongruence (i.e., not all genes follow the species history). 1 , 6 , 7 Making use of large amounts of genome data comes with both an important caveat and an important boon: while methodologically incongruent genes can be removed by a number of tools to identify branch length heterogeneity, compositional heterogeneity and site saturation, genes that are excluded due to biological incongruence may contain real biological information that alters our understanding of species relationships. 1 Currently, phylogenetics has adopted a conservative stance, 1 , 18 selecting genes that fit well within our models of evolution 19 , 20 – this has the benefit of evading artifactual topologies, but it may also be presenting us with hypotheses of evolution that are preselected according to our own biases. As a result, this could cause us to incorrectly adopt great confidence in hypotheses that are not as well supported by the data as they first appear. 21 In this respect, an approach that is conservative towards the models may not be conservative towards confidence in our phylogenetic hypotheses. Scoutknife presents an alternative. 11 , 22 By randomly sampling data across the genome, the key hurdle for this methodology is assessing whether methodological incongruence significantly negatively influences the final hypothesis, or whether the false signal supplied by these model violations is outweighed by the addition of the real biological signal supplied by the sheer density of big data. If the latter is true, Scoutknife may represent a better way forward for generating phylogenetic topologies – one that is robust to methodological incongruence whilst expressing the biological incongruence that is present in the data. Here we present Scoutknife, a new method for assessing topological support while simultaneously reconstructing phylogenetic relationships. In contrast to the traditional bootstrap and jackknifing approaches, Scoutknife discards the creation of artificial pseudo-replicates of sites to instead use large multi-gene inputs to create true replicate samples of a larger pool of genes. Through this procedure, Scoutknife is similar to gene jackknifing. Jackknifing of gene data has been historically used in two ways. First, to assess the effect of gene order on topological confidence in concatenated gene datasets. 23 Second, to reconstruct the phylogeny of a larger dataset by subsampling a random set of genes (or gene trees), 24 to assess the robustness of the obtained phylogeny, as in a traditional bootstrap approach. 25 – 27 Both jackknife and Scoutknife datasets are formed by reducing the overall number of genes in comparison to the original sample and the same gene will never occur twice in the same replicate. The first gene jackknifing procedure reduces the dataset via a subtractive process. 23 Jackknife in this form is a useful tool for understanding the impact of gene order on topologies - by otherwise keeping order constant, the effect of removal can be directly assessed. 23 In contrast, the second form for gene jackknifing and Scoutknife draw their samples randomly from the source dataset. The difference between gene jackknifing as applied nowadays 25 , 26 and Scoutknife lies primarily in the subtle difference in their philosophical basis. The second form of gene jackknifing is applied after the phylogenetic reconstruction based on the entire dataset and with the sole purpose of assessing the robustness of the results ( Figure 1C ). In contrast, Scoutknife is a naive and unbiased way to measure support with genome-scale data, while reconstructing phylogenetic relationships. It does this by generating a sample number of datasets, each consisting of a user-specified number of randomly selected genes and forming a consensus tree from the results ( Figure 1 ). Scoutknife can also be used to implement the a posteriori gene jackknifing procedure. To our knowledge, it is the first user-friendly implementation of this procedure. Scoutknife support values can be attached to nodes of a maximum likelihood tree, similar to conventional bootstrap or jackknife support. Here we show, across 18 real and 100 simulation datasets, Scoutknife consensus trees produce comparable topological results to selecting the best genes within the dataset using GeneSortR, 19 and is robust to poor data occupancy. In addition, Scoutknife proves to be more granular in its assessment of topological reliability than traditional bootstrap values, allowing researchers to be more cautious and informed about their topological hypotheses than ever before. Approach Scoutknife takes a “brute force” approach to assessing phylogenetic robustness, simply asking the question - “how robust is the most likely tree to topological signal across the entire dataset?”. Rather than generating pseudo-samples by randomly sampling sites, as in a traditional bootstrap, 11 Scoutknife generates real data samples by randomly sampling genes to create randomly assembled concatenated multi-gene datasets ( Figure 1C ). In theory, though some of these genes contain low signal and others contain signal not consistent with the species phylogeny – either by methodological or biological incongruence 1 , 7 – the majority should contain at least some signal of the overall species tree, thereby allowing us to more robustly quantify not only the degree of support for a given taxonomic topology, but also the degree of discordance within the constituent genomes themselves. This naïve method may further allow us to resolve new phylogenetic hypotheses that have previously been neglected due to a focus on data selection. First, the multi-gene dataset is divided into individual gene alignments. These alignments are then randomly selected to form 100 multi-gene partitioned datasets of a size equal to that selected by the user. The same gene cannot be selected twice by the same dataset ( Figure 1C ) – a key difference from a traditional bootstrap 11 – but may appear multiple times across different datasets. Within our real dataset analyses, this sampling comprised 100 100-gene datasets selected from multi-gene datasets ranging from 1049 to 5105 genes ( Table 1 ). Our simulated datasets comprised 100 replicates of a 1049 gene dataset, from which 100 100-gene datasets were then sampled. Materials & Methods Dataset construction and analysis For both real and simulation analyses (for details see below), 100 genes were randomly selected 100 times from the source datasets, generating 100 100-gene concatenated sample datasets using the Scoutknife Package ( https://github.com/JFFleming/Scoutknife ). The Scoutknife script package requires catsequences 28 to be installed as a prerequisite, available at ( https://github.com/ChrisCreevey/catsequences ). Phylogenies for each Scoutknife dataset were constructed under IQ-Tree v1.6.12 29 using ModelFinder, 30 with a separate model applied to each gene and no partition merging. As a data density-based technique, Scoutknife might be expected to perform better in high data density scenarios where partitions can be comfortably merged. As such, this was intended to limit the efficacy of Scoutknife further and test its performance under a scenario with more highly variable best fit models than might be expected under normal conditions, whilst conserving computational effort considering the large number of test datasets and simulations. To further facilitate parallelization of the analyses, the phylogenetic analyses of the datasets were submitted using Scoutknifette ( https://github.com/Togtja/scoutknifette ). Scoutknifette is a custom high-performance computing (HPC) webhook for the group messaging service Discord 31 that can be easily modified for any HPC tasks that require multiple submission batches and queue tracking. The trees produced by each Scoutknife sample dataset were then concatenated into a single treelist file (see Underlying Data in our Data Availability Statement), and a consensus tree was constructed using bpcomp, available in Phylobayes, 32 by using a burnin of 0 and a sampling rate of 1, sampling each tree in the treelist. Trees were constructed as both 70% strict consensus and 50% majority consensus trees, and the results were compared. In two cases (Araneae and Lepidoptera), 30% plurality consensus trees were constructed using the same method, to further explore the data, as explained in the results and discussion section. In a single case (Actinopterygii), the low occupancy of two species in particular ( Muraenesox cinerus with 1 gene and Scomber scombrus with 15 genes across the entire dataset of 1105) meant that many of the Scoutknife samples did not contain representatives from these taxa. To address this, we used sumtrees.py v 4.5.2, part of the DendroPy package, 33 as it is capable of building consensus trees from tree lists containing a variable number of taxa. The Quartet Similarity, Quartet Divergence, Node Conflict, Node Agreement, Strict Joint Assertions, Semi-Strict Joint Assertions, Symmetric Difference, Marczewski-Steinhaus, Steel-Penny and Overall Similarity were measured with reference to the previously published topology of the 250 most informative genes of that dataset, as selected by GeneSortR. 19 In the case of the simulated datasets, the topology of the 250 most informative genes of the original source dataset, Milla et al., (2020), as selected by GeneSortR, 19 , 34 was used. These similarity metrics were calculated using the ‘Quartet’ Library available in R. 35 Real test datasets To assess the efficacy of Scoutknife, we examined 18 real data datasets, 25 , 34 , 36 – 51 those used in a similar benchmarking study by GeneSortR. 19 These datasets range from 1049 to 5105 genes and from 30 to 332 taxa in size, comprising studies of animals, plants and fungi ( Table 1 ). In contrast to the prior study, genes with less than 50% occupancy were not removed: Scoutknife should show decreased performance at low occupancy levels, as it relies on data density, and so this should give a clearer picture of how the methodology performs across a variety of real datasets. The resultant tree topologies were then compared to the topology recovered by analysing the most informative 250 genes, as determined by GeneSortR, 19 to assess whether the same topological hypothesis was resolved by the Scoutknife Consensus Tree. Table 1. A table listing the real data datasets used to benchmark the performance of Scoutknife. The first column names the taxa that form the ingroup of the phylogeny. The second names the original publication (although all source datasets are the same as those evaluated by Koch et al. (2021). 19 The third and fourth columns detail the number of taxa and the number of genes in the original alignment respectively. Taxa Study Number of taxa Number of genes Actinopterygii Hughes et al. (2018) 36 306 1105 Araneae Fernández et al. (2018) 37 168 2365 Aspergillacea Steenwyk et al. (2019) 38 93 1668 Blattodea Evangelista et al. (2019) 39 66 3235 Echinoidea Mongiardino Koch & Thompson (2021) 40 37 2356 Gnathostomata Irisarri et al. (2017) 25 100 4593 Heliozelidae Milla et al. (2020) 34 46 1049 Hemipteroids Johnson et al. (2018) 41 193 2395 Hexapoda Misof et al. (2014) 42 144 1478 Hymenoptera Peters et al. (2017) 43 174 3256 Lepidoptera Kawahara et al. (2019) 44 203 2098 Monilophytes Shen, Jin et al. (2018) 45 73 2391 Myriapoda Fernández et al. (2016) 46 51 2131 Opiliones Fernández et al. (2017) 47 67 1550 Phasmatodea Simon et al. (2019) 48 61 1097 Pseudoscorpiones Benavides et al. (2019) 49 48 2473 Saccharomycotina Shen, Opulente et al. (2018) 50 343 5105 Scorpiones Sharma et al. (2018) 51 43 1464 Simulation datasets To further assess the efficacy of Scoutknife, we generated 100 simulation datasets using the Alignment Mimic function of AliSim, as implemented in IQTree v2.2.0. 52 , 53 For this, 100 simulations were independently created for each gene in the Milla et al., (2020) 34 Heliozelidae dataset, as it represented a small-sized dataset of those within our real data study, at 1049 genes, and as such should have presented a challenge for Scoutknife. Furthermore, AliSim’s alignment mimic 52 allows us to generate alignment datasets that mimic real genes, complete with low occupancy and reasonable variations in alignment length. Alisim was implemented with the following command: iqtree2 – alisim − s − − num − alignments 100 For each set of 1049 simulated genes, 100 100-gene Scoutknife datasets were constructed, and then analysed using IQ Tree as with the real datasets. The Quartet Similarity, Quartet Divergence, Node Conflict, Node Agreement, Strict Joint Assertions, Semi-Strict Joint Assertions, Symmetric Difference, Marczewski-Steinhaus, Steel-Penny and Overall Similarity were then measured with reference to the previously published topology generated by analysing the 250 most informative genes of the Milla et al. (2020) dataset as selected by GeneSortR. 19 As each gene was simulated independently, it should in theory retain the topology of that initial single gene dataset, thereby replicating the discordance present in the original dataset. Furthermore, by directly comparing our random samples of simulated datasets to the most informative genes of the source dataset, this should disadvantage Scoutknife, as some of the simulated data may support a separate alternative topology to either the single gene or the real informative gene topology. Assessing the efficacy of Scoutknife For each dataset, we calculated a variety of quartet-based similarity metrics: the Quartet Divergence, 54 the proportion of nodes that did not conflict between trees, the proportion of nodes that explicitly agreed between trees, the proportion of strict and semi-strict joint assertions, 55 the symmetric difference between trees 56 and the Steel-Penny 57 and Marczewski-Steinhaus similarity metrics. 56 Concordance with the initial study’s topology was first measured by assessing the proportion of nodes that explicitly agreed between topologies and then the proportion of nodes that did not conflict with the recovered topology. This could then be further scrutinized using the Quartet Divergence and then the Marczewski-Steinhaus (MS) measurement, which compares the distinctly resolved quartets in common between both trees. The remaining quartet measurements are present in our Underlying Data, available at DataDryad. Robinson-Foulds (RF) distances were not used due to Scoutknife’s propensity to recover nodes with conservatively low amounts of support. Polytomies are known to bias RF distances as they rely on a completely resolved tree, and this would be incompatible with the Scoutknife approach, which explicitly favours polytomies as representations of incongruent signal in the genome. 58 Results & Discussion Real datasets Across our 18 real test datasets, on a majority consensus tree, Scoutknife only struggles to recover the topology initially recovered by the original study in two cases (indicated by an explicit agreement of nodes below 90%, Quartet Divergence greater than 5% or a Marczewski-Steinhaus below 0.9) ( Figure 2 ). In the Araneae, Scoutknife achieved an “explicit agreement” value of 81.10%, Marczewski-Steinhaus of 0.80, and quartet divergence of 9.87%, which prompted us to further examine the dataset. The average occupancy of the dataset, when including genes with below 50% occupancy, is 46%. Furthermore, only 97 of the 2366 genes in the dataset had an occupancy greater than 80% ( Figure 3 ). In this case, it appears that Scoutknife struggles with lower resolution data, and that large amounts of missing data may be a genuine challenge to the efficacy of the method. However, when assessed using the more liberal criteria of measuring the proportion of nodes that do not conflict with the published tree (which is a measure that accounts for the uncertainty expressed by polytomies), 99.17% of recovered nodes were found to not be in explicit conflict ( Figure 2 ). This suggests that 18% of this discordance is caused by a conservative assessment of support in the data considering its low occupancy, not by disagreement in inference. Figure 2. A dual bar chart showing proportion of non-conflicting nodes (in blue) and explicitly agreeing nodes (in orange) for each dataset. The two datasets discussed further in the text, Araneae and Lepidoptera, are highlighted in light blue (for non-conflict) and red (for explicit agreement) respectively. Figure 3. A violin plot showing the distribution of gene occupancy across the Araneae dataset by Fernández et al. (2018). 31 A large proportion of low occupancy genes may cause issues for Scoutknife resolution. The second dataset that appeared to struggle under the Scoutknife approach was the Lepidoptera dataset. Here, only 81.66% of nodes were found to explicitly agree with the published topology, and it produced an MS value of 0.82 and quartet divergence of 9.19%. As in the Araneae, we find that 99.95% of the nodes did not conflict with the published topology. However, the reasons for this discordance within the Lepidoptera are less clear. This dataset had the sixth highest occupancy of the real datasets (88.8%), many of which produced more well-resolved Scoutknife consensus trees. Furthermore, GeneSortR measured the “Usefulness” of the dataset as the third highest of the selected study sets (0.33, on a range from 0.14-0.48). 19 Changing the minimum consensus value to produce a Scoutknife tree from a majority consensus tree to a 30% plurality support tree increases the Marczewski-Steinhaus value to 0.93, decreases quartet divergence to 3.63% and increases the number of nodes found to explicitly agree with the published topology to 92.83% (Underlying Data). This suggests that the discordance within the Lepidoptera dataset may be a true biological property of the history of the group, and that the difference between the Scoutknife result and prior published results may be indicative of gene selection and analysis methods strongly favouring one of a series of genuine alternative hypotheses that Scoutknife prefers to represent as a polytomy. This assertion is particularly supported when contrasted against the Araneae dataset – there, changing the minimum consensus value to produce a 30% plurality support tree increases the Marczewski-Steinhaus value from 0.80 to only 0.87 (0.07 increase Araneae vs. 0.11 in Lepidoptera), decreases quarter divergence from 9.87% to 6.77% (3.1% decrease vs 5.56% in Lepidoptera), and increases the number of nodes that “explicitly agree” from 81.10% to 87.59% (6.49% increase vs. 11.17% in Lepidoptera), a much smaller overall change in comparison. In the opposite direction, on a stricter 70% consensus tree, Scoutknife achieves an average of 99.85% nodes not conflicting with the trees produced by GeneSortR, ranging from 100% to 99.16%. At this higher value, however, explicit agreement varies between 56.91% (in the Araneae) and 99.90%, with an average of 94.35% (or 96.56% if the Araneae are excluded). This is due to the innate conservatism of Scoutknife – as the consensus guideline is increased, it is more likely to favour collapsing more nodes into polytomies – the average decrease in Explicit Agreement with the GeneSortR tree between the majority consensus trees and the 0.7 consensus tree is 2.91%, with values ranging from 0% (Echinoidea) to 24.18% (Araneae). Simulation datasets Within our simulation datasets, Scoutknife consistently recovers topologies that are consistent with the GeneSortR tree – the 70% strict consensus simulation trees recover no conflicting nodes with the GeneSortR topology. However, across the 100 simulation consensus trees, not all explicitly agree with the nodes resolved by GeneSortR ( Figure 4 ). At 70% strict consensus, explicit agreement varied between 97.81% and 88.64% with an average of 92.85%. This represents the greater conservatism of Scoutknife as a method – across all analyses, it prefers to resolve as polytomies, rather than bifurcations, representing the discordance across the genes in the dataset. This is further confirmed by the Marczewski-Steinhaus similarity index, which is consistent with the explicit agreement values (varying from 0.89 to 0.98 with an average of 0.93), suggesting that the only difference between the Scoutknife and GeneSortR topologies is in the existence of polytomies. Figure 4. A violin plot showing the distribution of Marczewski-Steinhaus values between Scoutknife Consensus trees and the GeneSortR Most Informative 250 Genes Tree at both a 0.7 strict consensus and 0.5 majority consensus. Note the long tail on the Majority Marczewkski-Steinhaus violin, representing Simulation 20. Examining the simple majority consensus trees, requiring a consensus of only 50% of resolved gene trees to resolve the node and not 70%, two bifurcating topologies were produced that conflicted with the GeneSortR topology (Simulation 20 and Simulation 98), reducing the average nodal “Do Not Conflict” value from 100% to 99.92%. While the Simulation 98 topology was very similar to the GeneSortR topology (Quartet Divergence 0.010), Simulation 20 showed significant divergence from the GeneSortR topology (Quarter Divergence 0.082). The discordance in Simulation 20 is caused by a single node distinguishing the Pseliastis group, Hoplophanes group and the ( Heliozela/Antispila/Antispilina/Holocacista/Coptodisca ) group. The Scoutknife analysis of Simulation 20 recovers this node with a support of 0.51 for ( Pseliastis + Hoplophanes ), the topology that is not favoured by GeneSortR or the remaining 99 Scoutknife simulations. GeneSortR recovered the alternative topology ( Pseliastis + Heliozela/Antispila/Antispilina/Holocacista/Coptodisca ) with a boostrap support of 83, the second least supported node in the entire Heliozelidae dataset, suggesting that there is considerable conflict at this node. The GeneSortR topology was also recovered by the Scoutknife analysis of the original dataset with a support of 0.62 and by the original study 34 with a UF bootstrap support of 65.1 and an SH-aLRT result of 72. Across our other Scoutknife simulations ( Pseliastis + Heliozela/Antispila/Antispilina/Holocacista/Coptodisca ), was recovered with support ranging from 0.57 to 0.86. As a particularly short branch in all analyses, this could suggest that Scoutknife struggles to discern the topology when fewer genes have the capacity to resolve a node, or when incomplete lineage sorting increases substantially due to short branch lengths. Across all 100 Simulation datasets, when the consensus value was lowered to a majority consensus tree, explicit agreement with the GeneSortR topology increased from an average of 92.85% to 98.62%, with explicit agreement values varying from 91.19% to 100%. This shows that, on average, in 5.77% of the nodes in the tree where GeneSortR displayed high confidence, Scoutknife instead assigned these nodes between 50 and 69% support. In accordance with this, Marczewski-Steinhaus similarity scores increased from an average of 0.93 to 0.99, with a variance from 0.85 – the aforementioned Simulation 20 – to 1 ( Figure 4 ). Discounting the outlying Simulation 20, Marczewski-Steinhaus similarity scores vary from only 0.97 to 1. This further showcases the benefits of Scoutknife’s more conservative approach, making use of the diversity of data to give a more informed approximation of support from the gene trees without necessarily losing resolution at these key nodes. Scoutknife and per-taxon gene occupancy: A hypergeometric distribution Our expectation was that as datasets became larger, they would become easier for Scoutknife to assess. However, instead, in both the Araneae and the simulation datasets, we found that Scoutknife was far more severely affected by per taxa dataset occupancy, rather than dataset size. In the simulation datasets, this takes the form of the simulation genes derived from Nothofagus , which is present in only 98 of the 1049 genes in each dataset. A consequence of this is that, in a truly representative 100-gene Scoutknife sample, a gene containing Nothofagus should be selected 9.3 times. As an individual Scoutknife sample cannot select the same gene twice (although the same gene can be selected multiple times between samples), the probability of selecting any given gene can be modeled as a hypergeometric distribution. This presents us with an understanding that only 60.52% of 100-gene Scoutknife samples will comprise at least 9 Nothofagus genes. On the other hand, there is a 99.99% chance that a 100-gene dataset contains at least one Nothofagus gene among the one hundred. However, in a 50-gene Scoutknife sample, there would be a 0.66% chance that 0 genes containing this taxon would be selected across the 1049. That means that across 100 50-gene Scoutknife samples, 1 sample of the 100 is likely to contain no representation of this taxon. In this way, taxa with low gene occupancy have a far more notable effect on Scoutknife than reducing the number of genes, which evenly reduces the number of genes for all taxa in the dataset. This is not surprising as the principal assumption of the ScoutKnife procedure ( Figure 1C ) is that given genome-scale data works as a surrogate for unlimited data ( Figure 1B ). Accordingly, the power of ScoutKnife is driven by the availability of genome-scale data across the entire dataset and not just parts of it. The taxa with the lowest genomic representation set the ceiling for Scoutknife’s effectiveness, rather than those with reference genomes. All the 18 datasets used for this study were compiled before the reference genome revolution, which is still very recent 16 , 17 and still restricted to only certain sections of the tree of life. Hence, for many taxa, genome-scale data at EBP minimum standards 59 are still lacking. However, in the near future, the full potential of ScoutKnife can be brought to bear on these data. Our analyses already strongly indicate the potential of these methods in comparison to others through their conservativism in tree resolution and support values due to a higher susceptibility to the biological and methodological incongruence in the data. In the meantime, the reduced power of ScoutKnife due to taxa with reduced genomic representation can be addressed by increasing the number of genes selected by a Scoutknife sample relative to poor taxon occupancy. This increases the absolute number of genes containing the low occupancy taxa in the dataset, though it will not affect the proportional representation of the low occupancy taxa. For example, to consider the Nothofagus earlier, a 200-gene Scoutknife dataset would increase the chance of observing 9 Nothofagus genes in any given Scoutknife sample from 60.52% to 99.84%. By doubling the size of the Scoutknife sample, a representative number of genes would be 18. However, simply increasing the raw representation of genes may aid Scoutknife resolution. This approach deviates from the naïve sampling strategy and introduces missingness as a selection parameter. On the other hand, this is often already done explicitly or implicitly as some genes can only be found in certain ingroups, for example, due to a gene duplication event, and so are generally excluded from these analyses in the dataset compilation step. Among the tools available at the Scoutknife Github is a Hypergeometric distribution calculator designed with Scoutknife in mind, to help researchers understand the composition of their Scoutknife samples prior to analysis. In addition, the Scoutknife “auto” function uses this distribution to calculate how many genes must be sampled across the input dataset in order to ensure a 99.9% chance of any given Scoutknife dataset containing that gene, and then automatically formats a Scoutknife analysis for the user based on these parameters. Conclusions Selection-based metrics have rightly dominated phylogenetic discussions for a great number of years, but in the era of big data, transitioning towards methods that make best use of the increased analytical power of whole genomes may be more prudent. Our results, and the Scoutknife methodology, show that, contrary to accepted wisdom, model violations and incongruence can be overcome by sheer density of data. What results is a more neutral look at phylogenetic relationships, rather than one biased by our own notions of what makes genes suitable for phylogenetics. A helpful side effect of this is an increase in computational efficiency: rather than assessing individual gene trees prior to multi-gene analysis, 100 smaller Scoutknife datasets assess the robustness of a total dataset analysis or form the basis of a consensus tree. In many cases across our datasets, Scoutknife appears to recover the same relationships as before, but is also able to quantify our confidence in hypotheses of shared evolution efficiently and conservatively. In the future, this may be critical to a more holistic view of phylogeny. In addition, as models improve, and model incongruence becomes less and less of a concern, as a model-neutral methodology, Scoutknife’s ability to assess true biological incongruence will only improve, making it not only an exciting option for the present, but an even more effective one in the future. Data availability Source data • Source Data sets, 25 , 34 , 36 – 51 and their analysis within GeneSortR 19 were used to assess the efficacy of Scoutknife. All files used to assess the efficacy of Scoutknife can be found reproduced in our underlying data link (below). Further information on the Source datasets can also be found in the supplemental data for Koch et al. (2021). 19 Underlying data • Both our real and simulated data analyses are available at DataDryad, along with copies of individual gene fasta files from Source Data sets 25 , 34 , 36 – 51 and the 250 most informative gene trees from Koch et al. (2021) 19 that were used to benchmark Scoutknife’s performance ( https://datadryad.org/stash/dataset/doi:10.5061/dryad.sxksn0383 ). Software availability • Scoutknife is available at GitHub https://github.com/JFFleming/Scoutknife . • Archived Scoutknife code is available at: 10.5281/zenodo.8160834 Data is available under the terms of a GNU General Public License v2.0 only Acknowledgements The authors would like to recognise Tomas Berger for developing Scoutknifette, making the large number of MPI submissions necessary for the project more manageable. References 1. Fleming JF, Valero-Gracia A, Struck TH: Identifying and addressing methodological incongruence in phylogenomics: A review. Evol. Appl. 2023; 16 : 1087–1104. PubMed Abstract | Publisher Full Text | Free Full Text 2. Wolfe KH, Li W-H: Molecular evolution meets the genomics revolution. Nat. Genet. 2003; 33 (3): 255–265. PubMed Abstract | Publisher Full Text 3. Gee H: Ending incongruence. Nature. 2003 2003/10; 425 (6960): 782. Publisher Full Text 4. Bortoluzzi C, Wright CJ, Lee S, et al. : Lepidoptera genomics based on 88 chromosomal reference sequences informs population genetic parameters for conservation. bioRxiv. 2023:2023.04.14.536868. 5. Challis R, Kumar S, Sotero-Caio C, et al. : Genomes on a Tree (GoaT): A versatile, scalable search engine for genomic and sequencing project metadata across the eukaryotic tree of life. Wellcome Open Res. 2023; 8 (24): 24. PubMed Abstract | Publisher Full Text | Free Full Text 6. Mirarab S, Nakhleh L, Warnow T: Multispecies Coalescent: Theory and Applications in Phylogenetics. Annu. Rev. Ecol. Evol. Syst. 2021 2021/11/02; 52 (1): 247–268. Publisher Full Text 7. Mirarab S, Bayzid MS, Warnow T: Evaluating Summary Methods for Multilocus Species Tree Estimation in the Presence of Incomplete Lineage Sorting. Syst. Biol. 2014 2014/08/26; 65 (3): 366–380. PubMed Abstract | Publisher Full Text 8. Erixon P, Svennblad B, Britton T, et al. : Reliability of Bayesian posterior probabilities and bootstrap frequencies in phylogenetics. Syst. Biol. 2003; 52 (5): 665–673. PubMed Abstract | Publisher Full Text 9. Anisimova M, Gil M, Dufayard J-F, et al. : Survey of branch support methods demonstrates accuracy, power, and robustness of fast likelihood-based approximation schemes. Syst. Biol. 2011; 60 (5): 685–699. PubMed Abstract | Publisher Full Text | Free Full Text 10. Anisimova M, Gascuel O: Approximate Likelihood-Ratio Test for Branches: A Fast, Accurate, and Powerful Alternative. Syst. Biol. 2006 2006/08/01; 55 (4): 539–552. PubMed Abstract | Publisher Full Text 11. Felsenstein J: Confidence limits on phylogenies: an approach using the bootstrap. Evolution. 1985; 39 (4): 783–791. PubMed Abstract | Publisher Full Text 12. Siddall ME: Another monophyly index: revisiting the jackknife. Cladistics. 1995; 11 (1): 33–56. PubMed Abstract | Publisher Full Text 13. Soltis PS, Soltis DE: Applying the bootstrap in phylogeny reconstruction. Stat. Sci. 2003; 18 : 256–267. Publisher Full Text 14. Swofford DL: Phylogenic inference. Molecular systematic. 1996. 15. Paez S, Kraus RH, Shapiro B, et al. : Reference genomes for conservation. Science. 2022; 377 (6604): 364–366. Publisher Full Text 16. Lewin HA, Richards S, Lieberman Aiden E, et al. : The earth BioGenome project 2020: Starting the clock. National Acad Sciences. 2022; 119 : e2115635118. PubMed Abstract | Publisher Full Text | Free Full Text 17. Ebenezer TE, Muigai AW, Nouala S, et al. : Africa: sequence 100,000 species to safeguard biodiversity. Nature. 2022; 603 (7901): 388–392. PubMed Abstract | Publisher Full Text 18. Lozano-Fernandez J: A Practical Guide to Design and Assess a Phylogenomic Study. Genome Biol. Evol. 2022; 14 (9): evac129. eng. PubMed Abstract | Publisher Full Text | Free Full Text 19. Mongiardino KN: Phylogenomic subsampling and the search for phylogenetically reliable loci. Mol. Biol. Evol. 2021; 38 (9): 4025–4038. PubMed Abstract | Publisher Full Text | Free Full Text 20. Edwards SV: Phylogenomic subsampling: a brief review. Zool. Scr. 2016; 45 : 63–74. Publisher Full Text 21. Rabiee M, Sayyari E, Mirarab S: Multi-allele species reconstruction using ASTRAL. Mol. Phylogenet. Evol. 2019; 130 : 286–296. PubMed Abstract | Publisher Full Text 22. Hillis DM, Moritz C, Mable BK: Molecular systematics. Sinauer; 1996. 23. Shi J, Zhang Y, Luo H, et al. : Using jackknife to assess the quality of gene order phylogenies.BMC Bioinformatics.2010; 11 (1): 168. PubMed Abstract | Publisher Full Text | Free Full Text 24. Simmons MP, Sloan DB, Springer MS, et al. : Gene-wise resampling outperforms site-wise resampling in phylogenetic coalescence analyses. Mol. Phylogenet. Evol. 2019; 131 : 80–92. PubMed Abstract | Publisher Full Text 25. Irisarri I, Baurain D, Brinkmann H, et al. : Phylotranscriptomic consolidation of the jawed vertebrate timetree. Mol. Phylogenet. Evol. 2017; 1 (9): 1370–1378. PubMed Abstract | Publisher Full Text | Free Full Text 26. Delsuc F, Tsagkogeorga G, Lartillot N, et al. : Additional molecular support for the new chordate phylogeny. Genesis. 2008; 46 (11): 592–604. Publisher Full Text 27. Philippe H, Poustka AJ, Chiodin M, et al. : Mitigating anticipated effects of systematic errors supports sister-group relationship between Xenacoelomorpha and Ambulacraria. Curr. Biol. 2019; 29 (11): 1818–1826.e6. PubMed Abstract | Publisher Full Text 28. Creevey C, Weeks N: ChrisCreevey/catsequences: Version 1.3. Zenodo. 2021. 29. Nguyen L-T, Schmidt HA, von Haeseler A , et al. : IQ-TREE: a fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies. Mol. Biol. Evol. 2015; 32 (1): 268–274. PubMed Abstract | Publisher Full Text 30. Darriba D, Posada D, Kozlov AM, et al. : ModelTest-NG: A New and Scalable Tool for the Selection of DNA and Protein Evolutionary Models.Mol. Biol. Evol.2020; 37 (1): 291–294. PubMed Abstract | Publisher Full Text | Free Full Text 31. Inc. D. Discord. 2020. https://discord.com 32. Lartillot N, Lepage T, Blanquart S: PhyloBayes 3: a Bayesian software package for phylogenetic reconstruction and molecular dating. Bioinformatics. 2009; 25 (17): 2286–2288. PubMed Abstract | Publisher Full Text 33. Sukumaran J, Holder MT: DendroPy: a Python library for phylogenetic computing. Bioinformatics. 2010; 26 (12): 1569–1571. PubMed Abstract | Publisher Full Text 34. Milla L, Moussalli A, Wilcox SA, et al. : Phylotranscriptomics resolves phylogeny of the Heliozelidae (Adeloidea: Lepidoptera) and suggests a Late Cretaceous origin in Australia. Syst. Entomol. 2020; 45 (1): 128–143. Publisher Full Text 35. Rdpack R, Rcpp L: Package ‘Quartet’. Adv. Appl. Math. 2019; 7 : 309–343. 36. Hughes LC, Ortí G, Huang Y, et al. : Comprehensive phylogeny of ray-finned fishes (Actinopterygii) based on transcriptomic and genomic data. Proc. Natl. Acad. Sci. 2018; 115 (24): 6249–6254. PubMed Abstract | Publisher Full Text | Free Full Text 37. Fernández R, Kallal RJ, Dimitrov D, et al. : Phylogenomics, diversification dynamics, and comparative transcriptomics across the spider tree of life. Curr. Biol. 2018; 28 (9): 1489–1497.e5. Publisher Full Text 38. Steenwyk JL, Shen X-X, Lind AL, et al. : A robust phylogenomic time tree for biotechnologically and medically important fungi in the genera Aspergillus and Penicillium. MBio. 2019; 10 (4): e00925–e00919. Publisher Full Text 39. Evangelista DA, Wipfler B, Béthoux O, et al. : An integrative phylogenomic approach illuminates the evolutionary history of cockroaches and termites (Blattodea). Proc. R. Soc. B. 1895; 2019 (286): 20182076. 40. Mongiardino Koch N, Thompson JR: A total-evidence dated phylogeny of Echinoidea combining phylogenomic and paleontological data. Syst. Biol. 2021; 70 (3): 421–439. Publisher Full Text 41. Johnson KP, Dietrich CH, Friedrich F, et al. : Phylogenomics and the evolution of hemipteroid insects. Proc. Natl. Acad. Sci. 2018; 115 (50): 12775–12780. PubMed Abstract | Publisher Full Text | Free Full Text 42. Misof B, Liu S, Meusemann K, et al. : Phylogenomics resolves the timing and pattern of insect evolution. Science. 2014; 346 (6210): 763–767. PubMed Abstract | Publisher Full Text 43. Peters RS, Krogmann L, Mayer C, et al. : Evolutionary history of the Hymenoptera. Curr. Biol. 2017; 27 (7): 1013–1018. Publisher Full Text 44. Kawahara AY, Plotkin D, Espeland M, et al. : Phylogenomics reveals the evolutionary timing and pattern of butterflies and moths. Proc. Natl. Acad. Sci. 2019; 116 (45): 22657–22663. PubMed Abstract | Publisher Full Text | Free Full Text 45. Shen H, Jin D, Shu J-P, et al. : Large-scale phylogenomic analysis resolves a backbone phylogeny in ferns. GigaScience. 2018; 7 (2): gix116. Publisher Full Text 46. Fernández R, Edgecombe GD, Giribet G: Exploring phylogenetic relationships within Myriapoda and the effects of matrix composition and occupancy on phylogenomic reconstruction. Syst. Biol. 2016; 65 (5): 871–889. PubMed Abstract | Publisher Full Text | Free Full Text 47. Fernández R, Sharma PP, Tourinho AL, et al. : The Opiliones tree of life: shedding light on harvestmen relationships through transcriptomics. Proc. R. Soc. B Biol. Sci. 2017; 284 (1849): 20162340. PubMed Abstract | Publisher Full Text | Free Full Text 48. Simon S, Letsch H, Bank S, et al. : Old World and New World Phasmatodea: phylogenomics resolve the evolutionary history of stick and leaf insects. Front. Ecol. Evol. 2019; 7 : 345. Publisher Full Text 49. Benavides LR, Cosgrove JG, Harvey MS, et al. : Phylogenomic interrogation resolves the backbone of the Pseudoscorpiones tree of life. Mol. Phylogenet. Evol. 2019; 139 : 106509. Publisher Full Text 50. Shen X-X, Opulente DA, Kominek J, et al. : Tempo and mode of genome evolution in the budding yeast subphylum. Cell. 2018; 175 (6): 1533–1545.e20. PubMed Abstract | Publisher Full Text | Free Full Text 51. Sharma PP, Baker CM, Cosgrove JG, et al. : A revised dated phylogeny of scorpions: phylogenomic support for ancient divergence of the temperate Gondwanan family Bothriuridae. Mol. Phylogenet. Evol. 2018; 122 : 37–45. PubMed Abstract | Publisher Full Text 52. Ly-Trong N, Naser-Khdour S, Lanfear R, et al. : Alisim: A fast and versatile phylogenetic sequence simulator for the genomic era. Mol. Biol. Evol. 2022; 39 (5): msac092. PubMed Abstract | Publisher Full Text | Free Full Text 53. Minh BQ, Schmidt HA, Chernomor O, et al. : IQ-TREE 2: new models and efficient methods for phylogenetic inference in the genomic era. Mol. Biol. Evol. 2020; 37 (5): 1530–1534. PubMed Abstract | Publisher Full Text | Free Full Text 54. Smith MR: Bayesian and parsimony approaches reconstruct informative trees from simulated morphological datasets. Biol. Lett. 2019; 15 (2): 20180632. Publisher Full Text 55. Estabrook GF, McMorris F, Meacham CA: Comparison of undirected phylogenetic trees based on subtrees of four evolutionary units. Syst. Zool. 1985; 34 (2): 193–200. Publisher Full Text 56. Day WHE: Analysis of Quartet Dissimilarity Measures Between Undirected Phylogenetic Trees. Syst. Biol. 1986; 35 (3): 325–333. Publisher Full Text 57. Steel MA, Penny D: Distributions of tree comparison metrics—some new results. Syst. Biol. 1993; 42 (2): 126–141. 58. Simmons MP, Goloboff PA, Stöver BC, et al. : Quantification of congruence among gene trees with polytomies using overall success of resolution for phylogenomic coalescent analyses. Cladistics. 59. Project EB. Reference Source Comments on this article Comments (0) Version 2 VERSION 2 PUBLISHED 07 Aug 2023 ADD YOUR COMMENT Comment Author details Author details 1 Natural History Museum, Universitetet i Oslo, Oslo, Oslo, 0562, Norway James Fleming Roles: Conceptualization, Formal Analysis, Investigation, Methodology, Software, Writing – Original Draft Preparation Pia Merete Eriksen Roles: Formal Analysis, Investigation, Writing – Review & Editing Torsten Hugo Struck Roles: Conceptualization, Funding Acquisition, Methodology, Resources, Supervision, Writing – Review & Editing Competing interests No competing interests were disclosed. Grant information This work was funded by the Research Council Norway (project number 300587 to T.H.S.). T.H.S. received additional support by the Norwegian Metacenter for Computational Science (NOTUR; project numbers NN9408K, and NS9408K). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Article Versions (2) version 2 Revised Published: 10 Jul 2024, 12:945 https://doi.org/10.12688/f1000research.139356.2 version 1 Published: 07 Aug 2023, 12:945 https://doi.org/10.12688/f1000research.139356.1 Copyright © 2024 Fleming J et al . This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Download Export To Sciwheel Bibtex EndNote ProCite Ref. Manager (RIS) Sente metrics Views Downloads F1000Research - - PubMed Central info_outline Data from PMC are received and updated monthly. - - Citations open_in_new 0 open_in_new 0 open_in_new SEE MORE DETAILS CITE how to cite this article Fleming J, Eriksen PM and Struck TH. Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric [version 2; peer review: 3 approved] . F1000Research 2024, 12 :945 ( https://doi.org/10.12688/f1000research.139356.2 ) NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS track receive updates on this article Track an article to receive email alerts on any updates to this article. TRACK THIS ARTICLE Share Open Peer Review Current Reviewer Status: ? Key to Reviewer Statuses VIEW HIDE Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Version 2 VERSION 2 PUBLISHED 10 Jul 2024 Revised Views 0 Cite How to cite this report: Kan X. Reviewer Report For: Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric [version 2; peer review: 3 approved] . F1000Research 2024, 12 :945 ( https://doi.org/10.5256/f1000research.168838.r301251 ) The direct URL for this report is: https://f1000research.com/articles/12-945/v2#referee-response-301251 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 18 Jul 2024 Xianzhao Kan , Anhui Normal University, Wuhu, China Approved VIEWS 0 https://doi.org/10.5256/f1000research.168838.r301251 In the revised manuscript, the authors have made substantial modifications as ... Continue reading READ ALL In the revised manuscript, the authors have made substantial modifications as requested. As a reviewer, I am satisfied and recommend it for approval. Competing Interests: No competing interests were disclosed. Reviewer Expertise: Bioinformatics, Genomics, and Molecular evolution. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Kan X. Reviewer Report For: Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric [version 2; peer review: 3 approved] . F1000Research 2024, 12 :945 ( https://doi.org/10.5256/f1000research.168838.r301251 ) The direct URL for this report is: https://f1000research.com/articles/12-945/v2#referee-response-301251 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Respond or Comment COMMENT ON THIS REPORT Views 0 Cite How to cite this report: Zaharias P. Reviewer Report For: Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric [version 2; peer review: 3 approved] . F1000Research 2024, 12 :945 ( https://doi.org/10.5256/f1000research.168838.r301250 ) The direct URL for this report is: https://f1000research.com/articles/12-945/v2#referee-response-301250 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 11 Jul 2024 Paul Zaharias , Institut de Systématique, Evolution, Biodiversité (ISYEB UMR7205 – CNRS), Museum National d'Histoire Naturelle, Paris, Île-de-France, France Approved VIEWS 0 https://doi.org/10.5256/f1000research.168838.r301250 I am satisfied with this version and have no further ... Continue reading READ ALL I am satisfied with this version and have no further comments to make. Thanks to the authors and editorial team! Competing Interests: No competing interests were disclosed. Reviewer Expertise: Phylogenetics, systematics and bioinformatics I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Zaharias P. Reviewer Report For: Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric [version 2; peer review: 3 approved] . F1000Research 2024, 12 :945 ( https://doi.org/10.5256/f1000research.168838.r301250 ) The direct URL for this report is: https://f1000research.com/articles/12-945/v2#referee-response-301250 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Respond or Comment COMMENT ON THIS REPORT Version 1 VERSION 1 PUBLISHED 07 Aug 2023 Views 0 Cite How to cite this report: Redmond A. Reviewer Report For: Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric [version 2; peer review: 3 approved] . F1000Research 2024, 12 :945 ( https://doi.org/10.5256/f1000research.152626.r195414 ) The direct URL for this report is: https://f1000research.com/articles/12-945/v1#referee-response-195414 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 25 May 2024 Anthony K. Redmond , Smurfit Institute of Genetics, Trinity College Dublin, Dublin, Ireland Approved VIEWS 0 https://doi.org/10.5256/f1000research.152626.r195414 This well written and presented study by Fleming and colleagues provides an important and much needed alternative measure of robustness for phylogenetic inference based on gene-wise jackknife resampling. The logical and practical ideas behind, and advantages of using, their ... Continue reading READ ALL This well written and presented study by Fleming and colleagues provides an important and much needed alternative measure of robustness for phylogenetic inference based on gene-wise jackknife resampling. The logical and practical ideas behind, and advantages of using, their approach are clearly explained and exemplified in comparison to the traditional site-wise bootstrapping approach. While I must admit that the study has not entirely convinced me that 'model violations and incongruence can be overcome by sheer density of data' (and this may be a bias of my own), I am nonetheless very pleased with the conservative and nuanced results provided by the new Scoutknife approach as compared to standard bootstrap. I believe the Scoutknife approach has great potential within the field and am happy to recommend the manuscript without revision. I am excited to see how the approach holds up to extreme lineage-specific violations on the modelling and orthology fronts. As a very minor point and optional point, I believe there may be additional studies worthy of discussion here that support some of the arguements made in the text. For example, Simmons et al. 2019 [ ref 1 ] previously proposed the use of gene-wise bootstrap for summary coalescent species tree methods, while gene wise bootstrapping has been used in other studies in the past (e.g. Simion et al 2017 [ ref 2 ]), and off the top of my head has been suggested to be conservative in at least Philippe et al 2019 [ ref 3 ]. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes References 1. Simmons MP, Sloan DB, Springer MS, Gatesy J: Gene-wise resampling outperforms site-wise resampling in phylogenetic coalescence analyses. Mol Phylogenet Evol . 2019; 131 : 80-92 PubMed Abstract | Publisher Full Text 2. Simion P, Philippe H, Baurain D, Jager M, et al.: A Large and Consistent Phylogenomic Dataset Supports Sponges as the Sister Group to All Other Animals. Curr Biol . 2017; 27 (7): 958-967 PubMed Abstract | Publisher Full Text 3. Philippe H, Poustka AJ, Chiodin M, Hoff KJ, et al.: Mitigating Anticipated Effects of Systematic Errors Supports Sister-Group Relationship between Xenacoelomorpha and Ambulacraria. Curr Biol . 2019; 29 (11): 1818-1826.e6 PubMed Abstract | Publisher Full Text Competing Interests: No competing interests were disclosed. Reviewer Expertise: Phylogenomics, Genome Evolution, Gene and Genome Duplication, Immunology I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Redmond A. Reviewer Report For: Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric [version 2; peer review: 3 approved] . F1000Research 2024, 12 :945 ( https://doi.org/10.5256/f1000research.152626.r195414 ) The direct URL for this report is: https://f1000research.com/articles/12-945/v1#referee-response-195414 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Respond or Comment COMMENT ON THIS REPORT Views 0 Cite How to cite this report: Kan X. Reviewer Report For: Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric [version 2; peer review: 3 approved] . F1000Research 2024, 12 :945 ( https://doi.org/10.5256/f1000research.152626.r261535 ) The direct URL for this report is: https://f1000research.com/articles/12-945/v1#referee-response-261535 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 06 May 2024 Xianzhao Kan , Anhui Normal University, Wuhu, China Approved with Reservations VIEWS 0 https://doi.org/10.5256/f1000research.152626.r261535 The manuscript, "Scoutknife: A Naïve, Whole-Genome Informed Phylogenetic Robustness Metric", introduces a novel tool, Scoutknife. In this method, the authors can potentially abandon the notion of pseudo-sampling and instead randomly sample small subsets of genes from the thousands of genes ... Continue reading READ ALL The manuscript, "Scoutknife: A Naïve, Whole-Genome Informed Phylogenetic Robustness Metric", introduces a novel tool, Scoutknife. In this method, the authors can potentially abandon the notion of pseudo-sampling and instead randomly sample small subsets of genes from the thousands of genes in their analyses. Furthermore, the authors validated the effectiveness of Scoutknife by applying it to 18 previously published datasets and 100 simulation studies. I believe that the development of this method and software will facilitate deeper research in phylogenetics. The following weaknesses should be addressed: 1. In the Abstract section, the term "robusticity" is used frequently, but I think it is not a standard statistical term. "Robustness" would be more suitable. 2. In the Conclusions of the Abstract, "phylogenetic hypotheses" should be changed to "phylogenetic hypothesis" because it refers to a singular hypothesis. 3. In the paragraph 4 of Introduction, the sentence "Currently, phylogenetics has adopted a conservative stance, selecting genes that fit well within our models of evolution – this has the benefit of evading artifactual topologies, but may well be presenting us with hypotheses of evolution that are preselected according to our own biases, or incorrectly causing us to adopt great confidence in hypotheses that are not as well supported by the data as it first appears." is quite long and could be split into multiple sentences for clarity. 4. In the paragraph 6 of Introduction, the phrase "true replicate samples" may be confusing. It would be helpful to clarify the difference between "true replicates" and "artificial pseudo-replicates". 5. In the section on Scoutknife and per-taxon gene occupancy, the following sentence is unclear: "genome-scale data works as a surrogate for unlimited data". The authors should clarify the relationship between genome-scale data and the concept of unlimited data. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Partly If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes Competing Interests: No competing interests were disclosed. Reviewer Expertise: Bioinformatics, Genomics, and Molecular evolution. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Kan X. Reviewer Report For: Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric [version 2; peer review: 3 approved] . F1000Research 2024, 12 :945 ( https://doi.org/10.5256/f1000research.152626.r261535 ) The direct URL for this report is: https://f1000research.com/articles/12-945/v1#referee-response-261535 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Author Response 10 Jul 2024 James Fleming , Natural History Museum, Universitetet i Oslo, Oslo, 0562, Norway 10 Jul 2024 Author Response We thank the reviewer for their consideration of our manuscript, and have attempted to clarify and address our text. We would like to especially thank them for point out the ... Continue reading We thank the reviewer for their consideration of our manuscript, and have attempted to clarify and address our text. We would like to especially thank them for point out the lack of clarity in points 4 and 5: these are quite critical concepts to understanding the wider manuscript, so it is important they are explained as clearly as possible. In order to address this, we have greatly revised the initial introduction section in order to hopefully explain the concepts and reasoning behind Scoutknife more clearly. We thank the reviewer for their consideration of our manuscript, and have attempted to clarify and address our text. We would like to especially thank them for point out the lack of clarity in points 4 and 5: these are quite critical concepts to understanding the wider manuscript, so it is important they are explained as clearly as possible. In order to address this, we have greatly revised the initial introduction section in order to hopefully explain the concepts and reasoning behind Scoutknife more clearly. Competing Interests: No competing interests were disclosed. Close Report a concern Respond or Comment COMMENTS ON THIS REPORT Author Response 10 Jul 2024 James Fleming , Natural History Museum, Universitetet i Oslo, Oslo, 0562, Norway 10 Jul 2024 Author Response We thank the reviewer for their consideration of our manuscript, and have attempted to clarify and address our text. We would like to especially thank them for point out the ... Continue reading We thank the reviewer for their consideration of our manuscript, and have attempted to clarify and address our text. We would like to especially thank them for point out the lack of clarity in points 4 and 5: these are quite critical concepts to understanding the wider manuscript, so it is important they are explained as clearly as possible. In order to address this, we have greatly revised the initial introduction section in order to hopefully explain the concepts and reasoning behind Scoutknife more clearly. We thank the reviewer for their consideration of our manuscript, and have attempted to clarify and address our text. We would like to especially thank them for point out the lack of clarity in points 4 and 5: these are quite critical concepts to understanding the wider manuscript, so it is important they are explained as clearly as possible. In order to address this, we have greatly revised the initial introduction section in order to hopefully explain the concepts and reasoning behind Scoutknife more clearly. Competing Interests: No competing interests were disclosed. Close Report a concern COMMENT ON THIS REPORT Views 0 Cite How to cite this report: Zaharias P. Reviewer Report For: Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric [version 2; peer review: 3 approved] . F1000Research 2024, 12 :945 ( https://doi.org/10.5256/f1000research.152626.r227172 ) The direct URL for this report is: https://f1000research.com/articles/12-945/v1#referee-response-227172 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 29 Dec 2023 Paul Zaharias , Institut de Systématique, Evolution, Biodiversité (ISYEB UMR7205 – CNRS), Museum National d'Histoire Naturelle, Paris, Île-de-France, France Approved with Reservations VIEWS 0 https://doi.org/10.5256/f1000research.152626.r227172 This article introduces “Scoutknife”, a software designed for gene jackknifing in phylogenomic datasets. The authors evaluate their approach with numerous empirical and simulated datasets and using a range of similarity metrics. They conclude that Scoutknife is a reliable estimator of ... Continue reading READ ALL This article introduces “Scoutknife”, a software designed for gene jackknifing in phylogenomic datasets. The authors evaluate their approach with numerous empirical and simulated datasets and using a range of similarity metrics. They conclude that Scoutknife is a reliable estimator of the robustness of phylogenetic hypotheses. I find the article well-written and easy to follow, and that “Scoutknife” could be a useful user-friendly software for phylogenomists. I only have two major comments, followed by some minor comments hereafter. First, the method is not new (contrarily to what can be read in the Introduction, paragraph 6); the procedure of gene jackknifing exists since at least 15 years ago [1,2]. As such, I would like to see in the Introduction a paragraph which would be a summary of the literature on the use of gene jackknifing, its assessed qualities, and drawbacks. This is however, the first time (to my knowledge) that a user-friendly software is suggested, and that there is a proper benchmarking of it, and the authors should emphasize on those aspects. Second, as the author point out in the discussion, one drawback of gene jackknifing (as it is with site-based jackknifing) is the problem of poor taxon occupancy, unfortunately a common problem in phylogenomic datasets. To address that, the authors wrote a “Hypergeometric distribution calculator” script to help the users better understand the taxon occupancy composition prior to analysis. I think this script should be automated and included as an option in the main “Scoutknife” approach as follow: The Scoutknife script automatically detects the number of genes in the folder and also automatically detects how many of those genes contain the least represented taxa. Then, Scoutknife calculates how many genes should be sampled at least so that there is >99.9% chances that the least represented taxon appears in at least one gene. This value could be considered the default minimal value of gene sampling for the user to guarantee the presence of all taxa in the subsets. Making the use of this “Taxon Probability Calculator" more visible and user-friendly would be a great plus and a great input to the more arbitrary “100 genes” strategy fixed by the authors and would show an adaptiveness of the approach to each dataset. Some minor comments: Everywhere: “Pseudosampling” in Fig.1 caption is not spelled the same way as other places in the text (“Pseudo sampling”). Might I suggest as well to add a hyphen after all “pseudo” as in “pseudo-sampling”, “pseudo-samples” and “pseudo-replicates”? Discussion, penultimate paragraph: please expand EBP to Earth BioGenome Project Last comment, which is more of a curiosity : is there any particular reason you chose the synonym "robusticity' instead of the more traditional "robustness" present in the literature ? Is the work clearly and accurately presented and does it cite the current literature? Partly Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes References 1. Delsuc F, Tsagkogeorga G, Lartillot N, Philippe H: Additional molecular support for the new chordate phylogeny. Genesis . 2008; 46 (11): 592-604 PubMed Abstract | Publisher Full Text 2. Irisarri I, Baurain D, Brinkmann H, Delsuc F, et al.: Phylotranscriptomic consolidation of the jawed vertebrate timetree. Nat Ecol Evol . 2017; 1 (9): 1370-1378 PubMed Abstract | Publisher Full Text Competing Interests: No competing interests were disclosed. Reviewer Expertise: Phylogenetics, systematics and bioinformatics I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Zaharias P. Reviewer Report For: Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric [version 2; peer review: 3 approved] . F1000Research 2024, 12 :945 ( https://doi.org/10.5256/f1000research.152626.r227172 ) The direct URL for this report is: https://f1000research.com/articles/12-945/v1#referee-response-227172 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Author Response 10 Jul 2024 James Fleming , Natural History Museum, Universitetet i Oslo, Oslo, 0562, Norway 10 Jul 2024 Author Response We would like to thank the reviewer for their positive response to the paper, and we found the comments below incredibly useful in improving the manuscript (which we are currently ... Continue reading We would like to thank the reviewer for their positive response to the paper, and we found the comments below incredibly useful in improving the manuscript (which we are currently uploading). To address their concerns one-by-one below: ================== First, the method is not new (contrarily to what can be read in the Introduction, paragraph 6); the procedure of gene jackknifing exists since at least 15 years ago [1,2]. As such, I would like to see in the Introduction a paragraph which would be a summary of the literature on the use of gene jackknifing, its assessed qualities, and drawbacks. This is however, the first time (to my knowledge) that a user-friendly software is suggested, and that there is a proper benchmarking of it, and the authors should emphasize on those aspects. ----- Scoutknife is not quite the same as gene jacknifing. The two are similar but quite distinct in their mathematical underpinnings and philosophical approach - in particular, jackknifing as typically implemented preserves the relative order of genes in the genome, and is also used predominantly to assess the effect of order on the resultant dataset, following initial phylogenetic tree construction. Scoutknife is much more similar to a classical bootstrapping approach, where the creation of alignment sets does not respect gene order. However, the reviewer is totally correct that these are relatively minor distinctions between the two approaches that were not clearly delineated in our initial manuscript, and that leaving out discussion of gene jacknifing is an important oversight. We have revised the initial introduction section to discuss jacknifing, and the differences between scoutknife and jacknife in more detail. We have also been more computationally explicit about the intentions of Scoutknife, with a new script, Scoutknife_Consensus.pl, being included in the GitHub, to allow users to easily generate consensus trees within the package itself, rather than relying on alternative programs as we have done in the manuscript itself. ==================== Second, as the author point out in the discussion, one drawback of gene jackknifing (as it is with site-based jackknifing) is the problem of poor taxon occupancy, unfortunately a common problem in phylogenomic datasets. To address that, the authors wrote a “Hypergeometric distribution calculator” script to help the users better understand the taxon occupancy composition prior to analysis. I think this script should be automated and included as an option in the main “Scoutknife” approach as follow: The Scoutknife script automatically detects the number of genes in the folder and also automatically detects how many of those genes contain the least represented taxa. Then, Scoutknife calculates how many genes should be sampled at least so that there is >99.9% chances that the least represented taxon appears in at least one gene. This value could be considered the default minimal value of gene sampling for the user to guarantee the presence of all taxa in the subsets. Making the use of this “Taxon Probability Calculator" more visible and user-friendly would be a great plus and a great input to the more arbitrary “100 genes” strategy fixed by the authors and would show an adaptiveness of the approach to each dataset. ----- That is a great idea! We have included an "--auto" flag inside Scoutknife that works as the reviewer describes. ====================== Some minor comments: ------ We have made revisions based on all minor comments. For the reviewer's curiosity, it was mostly the first author writing naturally - I'm not sure I'd put too much thought into that particular terminology usage, but you are totally correct to point it out. I have adjusted the terminology there appropriately. We would like to thank the reviewer for their positive response to the paper, and we found the comments below incredibly useful in improving the manuscript (which we are currently uploading). To address their concerns one-by-one below: ================== First, the method is not new (contrarily to what can be read in the Introduction, paragraph 6); the procedure of gene jackknifing exists since at least 15 years ago [1,2]. As such, I would like to see in the Introduction a paragraph which would be a summary of the literature on the use of gene jackknifing, its assessed qualities, and drawbacks. This is however, the first time (to my knowledge) that a user-friendly software is suggested, and that there is a proper benchmarking of it, and the authors should emphasize on those aspects. ----- Scoutknife is not quite the same as gene jacknifing. The two are similar but quite distinct in their mathematical underpinnings and philosophical approach - in particular, jackknifing as typically implemented preserves the relative order of genes in the genome, and is also used predominantly to assess the effect of order on the resultant dataset, following initial phylogenetic tree construction. Scoutknife is much more similar to a classical bootstrapping approach, where the creation of alignment sets does not respect gene order. However, the reviewer is totally correct that these are relatively minor distinctions between the two approaches that were not clearly delineated in our initial manuscript, and that leaving out discussion of gene jacknifing is an important oversight. We have revised the initial introduction section to discuss jacknifing, and the differences between scoutknife and jacknife in more detail. We have also been more computationally explicit about the intentions of Scoutknife, with a new script, Scoutknife_Consensus.pl, being included in the GitHub, to allow users to easily generate consensus trees within the package itself, rather than relying on alternative programs as we have done in the manuscript itself. ==================== Second, as the author point out in the discussion, one drawback of gene jackknifing (as it is with site-based jackknifing) is the problem of poor taxon occupancy, unfortunately a common problem in phylogenomic datasets. To address that, the authors wrote a “Hypergeometric distribution calculator” script to help the users better understand the taxon occupancy composition prior to analysis. I think this script should be automated and included as an option in the main “Scoutknife” approach as follow: The Scoutknife script automatically detects the number of genes in the folder and also automatically detects how many of those genes contain the least represented taxa. Then, Scoutknife calculates how many genes should be sampled at least so that there is >99.9% chances that the least represented taxon appears in at least one gene. This value could be considered the default minimal value of gene sampling for the user to guarantee the presence of all taxa in the subsets. Making the use of this “Taxon Probability Calculator" more visible and user-friendly would be a great plus and a great input to the more arbitrary “100 genes” strategy fixed by the authors and would show an adaptiveness of the approach to each dataset. ----- That is a great idea! We have included an "--auto" flag inside Scoutknife that works as the reviewer describes. ====================== Some minor comments: ------ We have made revisions based on all minor comments. For the reviewer's curiosity, it was mostly the first author writing naturally - I'm not sure I'd put too much thought into that particular terminology usage, but you are totally correct to point it out. I have adjusted the terminology there appropriately. Competing Interests: No competing interests were disclosed. Close Report a concern Respond or Comment COMMENTS ON THIS REPORT Author Response 10 Jul 2024 James Fleming , Natural History Museum, Universitetet i Oslo, Oslo, 0562, Norway 10 Jul 2024 Author Response We would like to thank the reviewer for their positive response to the paper, and we found the comments below incredibly useful in improving the manuscript (which we are currently ... Continue reading We would like to thank the reviewer for their positive response to the paper, and we found the comments below incredibly useful in improving the manuscript (which we are currently uploading). To address their concerns one-by-one below: ================== First, the method is not new (contrarily to what can be read in the Introduction, paragraph 6); the procedure of gene jackknifing exists since at least 15 years ago [1,2]. As such, I would like to see in the Introduction a paragraph which would be a summary of the literature on the use of gene jackknifing, its assessed qualities, and drawbacks. This is however, the first time (to my knowledge) that a user-friendly software is suggested, and that there is a proper benchmarking of it, and the authors should emphasize on those aspects. ----- Scoutknife is not quite the same as gene jacknifing. The two are similar but quite distinct in their mathematical underpinnings and philosophical approach - in particular, jackknifing as typically implemented preserves the relative order of genes in the genome, and is also used predominantly to assess the effect of order on the resultant dataset, following initial phylogenetic tree construction. Scoutknife is much more similar to a classical bootstrapping approach, where the creation of alignment sets does not respect gene order. However, the reviewer is totally correct that these are relatively minor distinctions between the two approaches that were not clearly delineated in our initial manuscript, and that leaving out discussion of gene jacknifing is an important oversight. We have revised the initial introduction section to discuss jacknifing, and the differences between scoutknife and jacknife in more detail. We have also been more computationally explicit about the intentions of Scoutknife, with a new script, Scoutknife_Consensus.pl, being included in the GitHub, to allow users to easily generate consensus trees within the package itself, rather than relying on alternative programs as we have done in the manuscript itself. ==================== Second, as the author point out in the discussion, one drawback of gene jackknifing (as it is with site-based jackknifing) is the problem of poor taxon occupancy, unfortunately a common problem in phylogenomic datasets. To address that, the authors wrote a “Hypergeometric distribution calculator” script to help the users better understand the taxon occupancy composition prior to analysis. I think this script should be automated and included as an option in the main “Scoutknife” approach as follow: The Scoutknife script automatically detects the number of genes in the folder and also automatically detects how many of those genes contain the least represented taxa. Then, Scoutknife calculates how many genes should be sampled at least so that there is >99.9% chances that the least represented taxon appears in at least one gene. This value could be considered the default minimal value of gene sampling for the user to guarantee the presence of all taxa in the subsets. Making the use of this “Taxon Probability Calculator" more visible and user-friendly would be a great plus and a great input to the more arbitrary “100 genes” strategy fixed by the authors and would show an adaptiveness of the approach to each dataset. ----- That is a great idea! We have included an "--auto" flag inside Scoutknife that works as the reviewer describes. ====================== Some minor comments: ------ We have made revisions based on all minor comments. For the reviewer's curiosity, it was mostly the first author writing naturally - I'm not sure I'd put too much thought into that particular terminology usage, but you are totally correct to point it out. I have adjusted the terminology there appropriately. We would like to thank the reviewer for their positive response to the paper, and we found the comments below incredibly useful in improving the manuscript (which we are currently uploading). To address their concerns one-by-one below: ================== First, the method is not new (contrarily to what can be read in the Introduction, paragraph 6); the procedure of gene jackknifing exists since at least 15 years ago [1,2]. As such, I would like to see in the Introduction a paragraph which would be a summary of the literature on the use of gene jackknifing, its assessed qualities, and drawbacks. This is however, the first time (to my knowledge) that a user-friendly software is suggested, and that there is a proper benchmarking of it, and the authors should emphasize on those aspects. ----- Scoutknife is not quite the same as gene jacknifing. The two are similar but quite distinct in their mathematical underpinnings and philosophical approach - in particular, jackknifing as typically implemented preserves the relative order of genes in the genome, and is also used predominantly to assess the effect of order on the resultant dataset, following initial phylogenetic tree construction. Scoutknife is much more similar to a classical bootstrapping approach, where the creation of alignment sets does not respect gene order. However, the reviewer is totally correct that these are relatively minor distinctions between the two approaches that were not clearly delineated in our initial manuscript, and that leaving out discussion of gene jacknifing is an important oversight. We have revised the initial introduction section to discuss jacknifing, and the differences between scoutknife and jacknife in more detail. We have also been more computationally explicit about the intentions of Scoutknife, with a new script, Scoutknife_Consensus.pl, being included in the GitHub, to allow users to easily generate consensus trees within the package itself, rather than relying on alternative programs as we have done in the manuscript itself. ==================== Second, as the author point out in the discussion, one drawback of gene jackknifing (as it is with site-based jackknifing) is the problem of poor taxon occupancy, unfortunately a common problem in phylogenomic datasets. To address that, the authors wrote a “Hypergeometric distribution calculator” script to help the users better understand the taxon occupancy composition prior to analysis. I think this script should be automated and included as an option in the main “Scoutknife” approach as follow: The Scoutknife script automatically detects the number of genes in the folder and also automatically detects how many of those genes contain the least represented taxa. Then, Scoutknife calculates how many genes should be sampled at least so that there is >99.9% chances that the least represented taxon appears in at least one gene. This value could be considered the default minimal value of gene sampling for the user to guarantee the presence of all taxa in the subsets. Making the use of this “Taxon Probability Calculator" more visible and user-friendly would be a great plus and a great input to the more arbitrary “100 genes” strategy fixed by the authors and would show an adaptiveness of the approach to each dataset. ----- That is a great idea! We have included an "--auto" flag inside Scoutknife that works as the reviewer describes. ====================== Some minor comments: ------ We have made revisions based on all minor comments. For the reviewer's curiosity, it was mostly the first author writing naturally - I'm not sure I'd put too much thought into that particular terminology usage, but you are totally correct to point it out. I have adjusted the terminology there appropriately. Competing Interests: No competing interests were disclosed. Close Report a concern COMMENT ON THIS REPORT Comments on this article Comments (0) Version 2 VERSION 2 PUBLISHED 07 Aug 2023 ADD YOUR COMMENT Comment keyboard_arrow_left keyboard_arrow_right Open Peer Review Reviewer Status info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Reviewer Reports Invited Reviewers 1 2 3 Version 2 (revision) 10 Jul 24 read read Version 1 07 Aug 23 read read read Paul Zaharias , Museum National d'Histoire Naturelle, Paris, France Xianzhao Kan , Anhui Normal University, Wuhu, China Anthony K. Redmond , Trinity College Dublin, Dublin, Ireland Comments on this article All Comments (0) Add a comment Sign up for content alerts Sign Up You are now signed up to receive this alert Browse by related subjects keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2024 Kan X. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 18 Jul 2024 | for Version 2 Xianzhao Kan , Anhui Normal University, Wuhu, China 0 Views copyright © 2024 Kan X. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (0) Approved info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions In the revised manuscript, the authors have made substantial modifications as requested. As a reviewer, I am satisfied and recommend it for approval. Competing Interests No competing interests were disclosed. Reviewer Expertise Bioinformatics, Genomics, and Molecular evolution. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. reply Respond to this report Responses (0) Kan X. Peer Review Report For: Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric [version 2; peer review: 3 approved] . F1000Research 2024, 12 :945 ( https://doi.org/10.5256/f1000research.168838.r301251) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/12-945/v2#referee-response-301251 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2024 Zaharias P. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 11 Jul 2024 | for Version 2 Paul Zaharias , Institut de Systématique, Evolution, Biodiversité (ISYEB UMR7205 – CNRS), Museum National d'Histoire Naturelle, Paris, Île-de-France, France 0 Views copyright © 2024 Zaharias P. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (0) Approved info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions I am satisfied with this version and have no further comments to make. Thanks to the authors and editorial team! Competing Interests No competing interests were disclosed. Reviewer Expertise Phylogenetics, systematics and bioinformatics I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. reply Respond to this report Responses (0) Zaharias P. Peer Review Report For: Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric [version 2; peer review: 3 approved] . F1000Research 2024, 12 :945 ( https://doi.org/10.5256/f1000research.168838.r301250) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/12-945/v2#referee-response-301250 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2024 Redmond A. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 25 May 2024 | for Version 1 Anthony K. Redmond , Smurfit Institute of Genetics, Trinity College Dublin, Dublin, Ireland 0 Views copyright © 2024 Redmond A. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (0) Approved info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions This well written and presented study by Fleming and colleagues provides an important and much needed alternative measure of robustness for phylogenetic inference based on gene-wise jackknife resampling. The logical and practical ideas behind, and advantages of using, their approach are clearly explained and exemplified in comparison to the traditional site-wise bootstrapping approach. While I must admit that the study has not entirely convinced me that 'model violations and incongruence can be overcome by sheer density of data' (and this may be a bias of my own), I am nonetheless very pleased with the conservative and nuanced results provided by the new Scoutknife approach as compared to standard bootstrap. I believe the Scoutknife approach has great potential within the field and am happy to recommend the manuscript without revision. I am excited to see how the approach holds up to extreme lineage-specific violations on the modelling and orthology fronts. As a very minor point and optional point, I believe there may be additional studies worthy of discussion here that support some of the arguements made in the text. For example, Simmons et al. 2019 [ ref 1 ] previously proposed the use of gene-wise bootstrap for summary coalescent species tree methods, while gene wise bootstrapping has been used in other studies in the past (e.g. Simion et al 2017 [ ref 2 ]), and off the top of my head has been suggested to be conservative in at least Philippe et al 2019 [ ref 3 ]. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes References 1. Simmons MP, Sloan DB, Springer MS, Gatesy J: Gene-wise resampling outperforms site-wise resampling in phylogenetic coalescence analyses. Mol Phylogenet Evol . 2019; 131 : 80-92 PubMed Abstract | Publisher Full Text 2. Simion P, Philippe H, Baurain D, Jager M, et al.: A Large and Consistent Phylogenomic Dataset Supports Sponges as the Sister Group to All Other Animals. Curr Biol . 2017; 27 (7): 958-967 PubMed Abstract | Publisher Full Text 3. Philippe H, Poustka AJ, Chiodin M, Hoff KJ, et al.: Mitigating Anticipated Effects of Systematic Errors Supports Sister-Group Relationship between Xenacoelomorpha and Ambulacraria. Curr Biol . 2019; 29 (11): 1818-1826.e6 PubMed Abstract | Publisher Full Text Competing Interests No competing interests were disclosed. Reviewer Expertise Phylogenomics, Genome Evolution, Gene and Genome Duplication, Immunology I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. reply Respond to this report Responses (0) Redmond A. Peer Review Report For: Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric [version 2; peer review: 3 approved] . F1000Research 2024, 12 :945 ( https://doi.org/10.5256/f1000research.152626.r195414) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/12-945/v1#referee-response-195414 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2024 Kan X. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 06 May 2024 | for Version 1 Xianzhao Kan , Anhui Normal University, Wuhu, China 0 Views copyright © 2024 Kan X. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (1) Approved With Reservations info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions The manuscript, "Scoutknife: A Naïve, Whole-Genome Informed Phylogenetic Robustness Metric", introduces a novel tool, Scoutknife. In this method, the authors can potentially abandon the notion of pseudo-sampling and instead randomly sample small subsets of genes from the thousands of genes in their analyses. Furthermore, the authors validated the effectiveness of Scoutknife by applying it to 18 previously published datasets and 100 simulation studies. I believe that the development of this method and software will facilitate deeper research in phylogenetics. The following weaknesses should be addressed: 1. In the Abstract section, the term "robusticity" is used frequently, but I think it is not a standard statistical term. "Robustness" would be more suitable. 2. In the Conclusions of the Abstract, "phylogenetic hypotheses" should be changed to "phylogenetic hypothesis" because it refers to a singular hypothesis. 3. In the paragraph 4 of Introduction, the sentence "Currently, phylogenetics has adopted a conservative stance, selecting genes that fit well within our models of evolution – this has the benefit of evading artifactual topologies, but may well be presenting us with hypotheses of evolution that are preselected according to our own biases, or incorrectly causing us to adopt great confidence in hypotheses that are not as well supported by the data as it first appears." is quite long and could be split into multiple sentences for clarity. 4. In the paragraph 6 of Introduction, the phrase "true replicate samples" may be confusing. It would be helpful to clarify the difference between "true replicates" and "artificial pseudo-replicates". 5. In the section on Scoutknife and per-taxon gene occupancy, the following sentence is unclear: "genome-scale data works as a surrogate for unlimited data". The authors should clarify the relationship between genome-scale data and the concept of unlimited data. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Partly If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes Competing Interests No competing interests were disclosed. Reviewer Expertise Bioinformatics, Genomics, and Molecular evolution. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. reply Respond to this report Responses (1) Author Response 10 Jul 2024 James Fleming, Natural History Museum, Universitetet i Oslo, Oslo, 0562, Norway We thank the reviewer for their consideration of our manuscript, and have attempted to clarify and address our text. We would like to especially thank them for point out the lack of clarity in points 4 and 5: these are quite critical concepts to understanding the wider manuscript, so it is important they are explained as clearly as possible. In order to address this, we have greatly revised the initial introduction section in order to hopefully explain the concepts and reasoning behind Scoutknife more clearly. View more View less Competing Interests No competing interests were disclosed. reply Respond Report a concern Kan X. Peer Review Report For: Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric [version 2; peer review: 3 approved] . F1000Research 2024, 12 :945 ( https://doi.org/10.5256/f1000research.152626.r261535) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/12-945/v1#referee-response-261535 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2023 Zaharias P. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 29 Dec 2023 | for Version 1 Paul Zaharias , Institut de Systématique, Evolution, Biodiversité (ISYEB UMR7205 – CNRS), Museum National d'Histoire Naturelle, Paris, Île-de-France, France 0 Views copyright © 2023 Zaharias P. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (1) Approved With Reservations info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions This article introduces “Scoutknife”, a software designed for gene jackknifing in phylogenomic datasets. The authors evaluate their approach with numerous empirical and simulated datasets and using a range of similarity metrics. They conclude that Scoutknife is a reliable estimator of the robustness of phylogenetic hypotheses. I find the article well-written and easy to follow, and that “Scoutknife” could be a useful user-friendly software for phylogenomists. I only have two major comments, followed by some minor comments hereafter. First, the method is not new (contrarily to what can be read in the Introduction, paragraph 6); the procedure of gene jackknifing exists since at least 15 years ago [1,2]. As such, I would like to see in the Introduction a paragraph which would be a summary of the literature on the use of gene jackknifing, its assessed qualities, and drawbacks. This is however, the first time (to my knowledge) that a user-friendly software is suggested, and that there is a proper benchmarking of it, and the authors should emphasize on those aspects. Second, as the author point out in the discussion, one drawback of gene jackknifing (as it is with site-based jackknifing) is the problem of poor taxon occupancy, unfortunately a common problem in phylogenomic datasets. To address that, the authors wrote a “Hypergeometric distribution calculator” script to help the users better understand the taxon occupancy composition prior to analysis. I think this script should be automated and included as an option in the main “Scoutknife” approach as follow: The Scoutknife script automatically detects the number of genes in the folder and also automatically detects how many of those genes contain the least represented taxa. Then, Scoutknife calculates how many genes should be sampled at least so that there is >99.9% chances that the least represented taxon appears in at least one gene. This value could be considered the default minimal value of gene sampling for the user to guarantee the presence of all taxa in the subsets. Making the use of this “Taxon Probability Calculator" more visible and user-friendly would be a great plus and a great input to the more arbitrary “100 genes” strategy fixed by the authors and would show an adaptiveness of the approach to each dataset. Some minor comments: Everywhere: “Pseudosampling” in Fig.1 caption is not spelled the same way as other places in the text (“Pseudo sampling”). Might I suggest as well to add a hyphen after all “pseudo” as in “pseudo-sampling”, “pseudo-samples” and “pseudo-replicates”? Discussion, penultimate paragraph: please expand EBP to Earth BioGenome Project Last comment, which is more of a curiosity : is there any particular reason you chose the synonym "robusticity' instead of the more traditional "robustness" present in the literature ? Is the work clearly and accurately presented and does it cite the current literature? Partly Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes References 1. Delsuc F, Tsagkogeorga G, Lartillot N, Philippe H: Additional molecular support for the new chordate phylogeny. Genesis . 2008; 46 (11): 592-604 PubMed Abstract | Publisher Full Text 2. Irisarri I, Baurain D, Brinkmann H, Delsuc F, et al.: Phylotranscriptomic consolidation of the jawed vertebrate timetree. Nat Ecol Evol . 2017; 1 (9): 1370-1378 PubMed Abstract | Publisher Full Text Competing Interests No competing interests were disclosed. Reviewer Expertise Phylogenetics, systematics and bioinformatics I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. reply Respond to this report Responses (1) Author Response 10 Jul 2024 James Fleming, Natural History Museum, Universitetet i Oslo, Oslo, 0562, Norway We would like to thank the reviewer for their positive response to the paper, and we found the comments below incredibly useful in improving the manuscript (which we are currently uploading). To address their concerns one-by-one below: ================== First, the method is not new (contrarily to what can be read in the Introduction, paragraph 6); the procedure of gene jackknifing exists since at least 15 years ago [1,2]. As such, I would like to see in the Introduction a paragraph which would be a summary of the literature on the use of gene jackknifing, its assessed qualities, and drawbacks. This is however, the first time (to my knowledge) that a user-friendly software is suggested, and that there is a proper benchmarking of it, and the authors should emphasize on those aspects. ----- Scoutknife is not quite the same as gene jacknifing. The two are similar but quite distinct in their mathematical underpinnings and philosophical approach - in particular, jackknifing as typically implemented preserves the relative order of genes in the genome, and is also used predominantly to assess the effect of order on the resultant dataset, following initial phylogenetic tree construction. Scoutknife is much more similar to a classical bootstrapping approach, where the creation of alignment sets does not respect gene order. However, the reviewer is totally correct that these are relatively minor distinctions between the two approaches that were not clearly delineated in our initial manuscript, and that leaving out discussion of gene jacknifing is an important oversight. We have revised the initial introduction section to discuss jacknifing, and the differences between scoutknife and jacknife in more detail. We have also been more computationally explicit about the intentions of Scoutknife, with a new script, Scoutknife_Consensus.pl, being included in the GitHub, to allow users to easily generate consensus trees within the package itself, rather than relying on alternative programs as we have done in the manuscript itself. ==================== Second, as the author point out in the discussion, one drawback of gene jackknifing (as it is with site-based jackknifing) is the problem of poor taxon occupancy, unfortunately a common problem in phylogenomic datasets. To address that, the authors wrote a “Hypergeometric distribution calculator” script to help the users better understand the taxon occupancy composition prior to analysis. I think this script should be automated and included as an option in the main “Scoutknife” approach as follow: The Scoutknife script automatically detects the number of genes in the folder and also automatically detects how many of those genes contain the least represented taxa. Then, Scoutknife calculates how many genes should be sampled at least so that there is >99.9% chances that the least represented taxon appears in at least one gene. This value could be considered the default minimal value of gene sampling for the user to guarantee the presence of all taxa in the subsets. Making the use of this “Taxon Probability Calculator" more visible and user-friendly would be a great plus and a great input to the more arbitrary “100 genes” strategy fixed by the authors and would show an adaptiveness of the approach to each dataset. ----- That is a great idea! We have included an "--auto" flag inside Scoutknife that works as the reviewer describes. ====================== Some minor comments: ------ We have made revisions based on all minor comments. For the reviewer's curiosity, it was mostly the first author writing naturally - I'm not sure I'd put too much thought into that particular terminology usage, but you are totally correct to point it out. I have adjusted the terminology there appropriately. View more View less Competing Interests No competing interests were disclosed. reply Respond Report a concern Zaharias P. Peer Review Report For: Scoutknife: A naïve, whole genome informed phylogenetic robusticity metric [version 2; peer review: 3 approved] . F1000Research 2024, 12 :945 ( https://doi.org/10.5256/f1000research.152626.r227172) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/12-945/v1#referee-response-227172 Alongside their report, reviewers assign a status to the article: Approved - the paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations - A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved - fundamental flaws in the paper seriously undermine the findings and conclusions Adjust parameters to alter display View on desktop for interactive features Includes Interactive Elements View on desktop for interactive features Competing Interests Policy Provide sufficient details of any financial or non-financial competing interests to enable users to assess whether your comments might lead a reasonable person to question your impartiality. Consider the following examples, but note that this is not an exhaustive list: Examples of 'Non-Financial Competing Interests' Within the past 4 years, you have held joint grants, published or collaborated with any of the authors of the selected paper. You have a close personal relationship (e.g. parent, spouse, sibling, or domestic partner) with any of the authors. You are a close professional associate of any of the authors (e.g. scientific mentor, recent student). You work at the same institute as any of the authors. You hope/expect to benefit (e.g. favour or employment) as a result of your submission. You are an Editor for the journal in which the article is published. Examples of 'Financial Competing Interests' You expect to receive, or in the past 4 years have received, any of the following from any commercial organisation that may gain financially from your submission: a salary, fees, funding, reimbursements. You expect to receive, or in the past 4 years have received, shared grant support or other funding with any of the authors. You hold, or are currently applying for, any patents or significant stocks/shares relating to the subject matter of the paper you are commenting on. Stay Updated Sign up for content alerts and receive a weekly or monthly email with all newly published articles Register with F1000Research Already registered? Sign in Not now, thanks close PLEASE NOTE If you are an AUTHOR of this article, please check that you signed in with the account associated with this article otherwise we cannot automatically identify your role as an author and your comment will be labelled as a “User Comment”. If you are a REVIEWER of this article, please check that you have signed in with the account associated with this article and then go to your account to submit your report, please do not post your review here. If you do not have access to your original account, please contact us . All commenters must hold a formal affiliation as per our Policies . The information that you give us will be displayed next to your comment. User comments must be in English, comprehensible and relevant to the article under discussion. We reserve the right to remove any comments that we consider to be inappropriate, offensive or otherwise in breach of the User Comment Terms and Conditions . Commenters must not use a comment for personal attacks. When criticisms of the article are based on unpublished data, the data should be made available. I accept the User Comment Terms and Conditions Please confirm that you accept the User Comment Terms and Conditions. Affiliation ✕ refresh Please enter your institution. Note: To add your institution or organisation, start typing the name and then select the correct name from the list. Where applicable, the name will appear in both the original language and in English. Do not paste in the name. If the name does not appear in the drop-down list, we will display the information you have entered. ✕ refresh Country/Region * USA UK Canada China France Germany Afghanistan Aland Islands Albania Algeria American Samoa Andorra Angola Anguilla Antarctica Antigua and Barbuda Argentina Armenia Aruba Australia Austria Azerbaijan Bahamas Bahrain Bangladesh Barbados Belarus Belgium Belize Benin Bermuda Bhutan Bolivia Bosnia and Herzegovina Botswana Bouvet Island Brazil British Indian Ocean Territory British Virgin Islands Brunei Bulgaria Burkina Faso Burundi Cambodia Cameroon Canada Cape Verde Cayman Islands Central African Republic Chad Chile China Christmas Island Cocos (Keeling) Islands Colombia Comoros Congo Cook Islands Costa Rica Cote d'Ivoire Croatia Cuba Cyprus Czech Republic Democratic Republic of the Congo Denmark Djibouti Dominica Dominican Republic Ecuador Egypt El Salvador Equatorial Guinea Eritrea Estonia Ethiopia Falkland Islands Faroe Islands Federated States of Micronesia Fiji Finland France French Guiana French Polynesia French Southern Territories Gabon Georgia Germany Ghana Gibraltar Greece Greenland Grenada Guadeloupe Guam Guatemala Guernsey Guinea Guinea-Bissau Guyana Haiti Heard Island and Mcdonald Islands Holy See (Vatican City State) Honduras Hong Kong Hungary Iceland India Indonesia Iran Iraq Ireland Israel Italy Jamaica Japan Jersey Jordan Kazakhstan Kenya Kiribati Kosovo (Serbia and Montenegro) Kuwait Kyrgyzstan Lao People's Democratic Republic Latvia Lebanon Lesotho Liberia Libya Liechtenstein Lithuania Luxembourg Macao Madagascar Malawi Malaysia Maldives Mali Malta Marshall Islands Martinique Mauritania Mauritius Mayotte Mexico Minor Outlying Islands of the United States Moldova Monaco Mongolia Montenegro Montserrat Morocco Mozambique Myanmar Namibia Nauru Nepal Netherlands Antilles New Caledonia New Zealand Nicaragua Niger Nigeria Niue Norfolk Island North Korea North Macedonia Northern Mariana Islands Norway Oman Pakistan Palau Palestinian Territory Panama Papua New Guinea Paraguay Peru Philippines Pitcairn Poland Portugal Puerto Rico Qatar Reunion Romania Russian Federation Rwanda Saint Helena Saint Kitts and Nevis Saint Lucia Saint Pierre and Miquelon Saint Vincent and the Grenadines Samoa San Marino Sao Tome and Principe Saudi Arabia Senegal Serbia Seychelles Sierra Leone Singapore Slovakia Slovenia Solomon Islands Somalia South Africa South Georgia and the South Sandwich Is South Korea South Sudan Spain Sri Lanka Sudan Suriname Svalbard and Jan Mayen Swaziland Sweden Switzerland Syria Taiwan Tajikistan Tanzania Thailand The Gambia The Netherlands Timor-Leste Togo Tokelau Tonga Trinidad and Tobago Tunisia Turkey Turkmenistan Turks and Caicos Islands Tuvalu UK USA Uganda Ukraine United Arab Emirates United States Virgin Islands Uruguay Uzbekistan Vanuatu Venezuela Vietnam Wallis and Futuna West Bank and Gaza Strip Western Sahara Yemen Zambia Zimbabwe Please select your country/region. You must enter a comment. Competing Interests Please disclose any competing interests that might be construed to influence your judgment of the article's or peer review report's validity or importance. Competing Interests Policy Provide sufficient details of any financial or non-financial competing interests to enable users to assess whether your comments might lead a reasonable person to question your impartiality. Consider the following examples, but note that this is not an exhaustive list: Examples of 'Non-Financial Competing Interests' Within the past 4 years, you have held joint grants, published or collaborated with any of the authors of the selected paper. You have a close personal relationship (e.g. parent, spouse, sibling, or domestic partner) with any of the authors. You are a close professional associate of any of the authors (e.g. scientific mentor, recent student). You work at the same institute as any of the authors. You hope/expect to benefit (e.g. favour or employment) as a result of your submission. You are an Editor for the journal in which the article is published. Examples of 'Financial Competing Interests' You expect to receive, or in the past 4 years have received, any of the following from any commercial organisation that may gain financially from your submission: a salary, fees, funding, reimbursements. You expect to receive, or in the past 4 years have received, shared grant support or other funding with any of the authors. You hold, or are currently applying for, any patents or significant stocks/shares relating to the subject matter of the paper you are commenting on. Please state your competing interests The comment has been saved. An error has occurred. Please try again. Cancel Post var lTitle = "Scoutknife: A na\u00EFve, whole genome informed...".replace("'", ''); var linkedInUrl = "http://www.linkedin.com/shareArticle?url=https://f1000research.com/articles/12-945/v2" + "&title=" + encodeURIComponent(lTitle) + "&summary=" + encodeURIComponent('Read the article by '); var deliciousUrl = "https://del.icio.us/post?url=https://f1000research.com/articles/12-945/v2&title=" + encodeURIComponent(lTitle); var redditUrl = "http://reddit.com/submit?url=https://f1000research.com/articles/12-945/v2" + "&title=" + encodeURIComponent(lTitle); linkedInUrl += encodeURIComponent('Fleming J et al.'); var offsetTop = /chrome/i.test( navigator.userAgent ) ? 4 : -10; var addthis_config = { ui_offset_top: offsetTop, services_compact : "facebook,twitter,www.linkedin.com,www.mendeley.com,reddit.com", services_expanded : "facebook,twitter,www.linkedin.com,www.mendeley.com,reddit.com", services_custom : [ { name: "LinkedIn", url: linkedInUrl, icon:"/img/icon/at_linkedin.svg" }, { name: "Mendeley", url: "http://www.mendeley.com/import/?url=https://f1000research.com/articles/12-945/v2/mendeley", icon:"/img/icon/at_mendeley.svg" }, { name: "Reddit", url: redditUrl, icon:"/img/icon/at_reddit.svg" }, ] }; var addthis_share = { url: "https://f1000research.com/articles/12-945", templates : { twitter : "Scoutknife: A na\u00EFve, whole genome informed phylogenetic.... Fleming J et al., published by " + "@F1000Research" + ", https://f1000research.com/articles/12-945/v2" } }; if (typeof(addthis) != "undefined"){ addthis.addEventListener('addthis.ready', checkCount); addthis.addEventListener('addthis.menu.share', checkCount); } $(".f1r-shares-twitter").attr("href", "https://twitter.com/intent/tweet?text=" + addthis_share.templates.twitter); $(".f1r-shares-facebook").attr("href", "https://www.facebook.com/sharer/sharer.php?u=" + addthis_share.url); $(".f1r-shares-linkedin").attr("href", addthis_config.services_custom[0].url); $(".f1r-shares-reddit").attr("href", addthis_config.services_custom[2].url); $(".f1r-shares-mendelay").attr("href", addthis_config.services_custom[1].url); function checkCount(){ setTimeout(function(){ $(".addthis_button_expanded").each(function(){ var count = $(this).text(); if (count !== "" && count != "0") $(this).removeClass("is-hidden"); else $(this).addClass("is-hidden"); }); }, 1000); } close How to cite this report {{reportCitation}} Cancel Copy Citation Details $(function(){R.ui.buttonDropdowns('.dropdown-for-downloads');}); $(function(){R.ui.toolbarDropdowns('.toolbar-dropdown-for-downloads');}); $.get("/articles/acj/139356/168838") new F1000.Clipboard(); new F1000.ThesaurusTermsDisplay("articles", "article", "168838"); $(document).ready(function() { $( "#frame1" ).on('load', function() { var mydiv = $(this).contents().find("div"); var h = mydiv.height(); console.log(h) }); var tooltipLivingFigure = jQuery(".interactive-living-figure-label .icon-more-info"), titleLivingFigure = tooltipLivingFigure.attr("title"); tooltipLivingFigure.simpletip({ fixed: true, position: ["-115", "30"], baseClass: 'small-tooltip', content:titleLivingFigure + " " }); tooltipLivingFigure.removeAttr("title"); $("body").on("click", ".cite-living-figure", function(e) { e.preventDefault(); var ref = $(this).attr("data-ref"); $(this).closest(".living-figure-list-container").find("#" + ref).fadeIn(200); }); $("body").on("click", ".close-cite-living-figure", function(e) { e.preventDefault(); $(this).closest(".popup-window-wrapper").fadeOut(200); }); $(document).on("mouseup", function(e) { var metricsContainer = $(".article-metrics-popover-wrapper"); if (!metricsContainer.is(e.target) && metricsContainer.has(e.target).length === 0) { $(".article-metrics-close-button").click(); } }); var articleId = $('#articleId').val(); if($("#main-article-count-box").attachArticleMetrics) { $("#main-article-count-box").attachArticleMetrics(articleId, { articleMetricsView: true }); } }); var figshareWidget = $(".new_figshare_widget"); if (figshareWidget.length > 0) { window.figshare.load("f1000", function(Widget) { // Select a tag/tags defined in your page. In this tag we will place the widget. _.map(figshareWidget, function(el){ var widget = new Widget({ articleId: $(el).attr("figshare_articleId") //height:300 // this is the height of the viewer part. [Default: 550] }); widget.initialize(); // initialize the widget widget.mount(el); // mount it in a tag that's on your page // this will save the widget on the global scope for later use from // your JS scripts. This line is optional. //window.widget = widget; }); }); } close Error Close Add Reset F1000.MICROSERVICES.AFFILIATION = ''; $(document).ready(function () { $('.js-affiliations-form').each((index, form) => { new AffiliationForm({ formId: form.id, institutionErrorSelector: '.comment-enter-institution', departmentErrorSelector: '.comment-enter-department', placeSelector: '.js-add-comment-place', stateSelector: '.js-add-comment-state', zipCodeSelector: '.js-add-comment-zipcode', countrySelector: '.js-add-comment-country', countryErrorSelector: '.comment-enter-country', }); }); }); $(document).ready(function () { var reportIds = { "301252": 0, "301250": 3, "301251": 3, "195410": 0, "195411": 0, "195414": 7, "195415": 0, "195412": 0, "195413": 0, "261531": 0, "261530": 0, "195416": 0, "252127": 0, "261535": 14, "261534": 0, "252126": 0, "261533": 0, "252125": 0, "261532": 0, "252124": 0, "261539": 0, "252131": 0, "261538": 0, "252130": 0, "261537": 0, "252129": 0, "261536": 0, "252128": 0, "252133": 0, "227172": 21, "208356": 0, "252132": 0, "210924": 0, }; $(".referee-response-container,.js-referee-report").each(function(index, el) { var reportId = $(el).attr("data-reportid"), reportCount = reportIds[reportId] || 0; $(el).find(".comments-count-container,.js-referee-report-views").html(reportCount); }); var uuidInput = $("#article_uuid"), oldUUId = uuidInput.val(), newUUId = "7af2156a-4160-4556-beef-3a79215bdccd"; uuidInput.val(newUUId); $("a[href*='article_uuid=']").each(function(index, el) { var newHref = $(el).attr("href").replace(oldUUId, newUUId); $(el).attr("href", newHref); }); }); An innovative open access publishing platform offering rapid publication and open peer review, whilst supporting data deposition and sharing. Browse Gateways Collections How it Works Contact For Developers Cookie Notice Privacy Notice RSS Submit Your Research Follow us © 2012-2026 F1000 Research Ltd. ISSN 2046-1402 | Legal | Partner of Research4Life • CrossRef • ORCID • FAIRSharing R.templateTests.simpleTemplate = R.template(' $text $text $text $text $text '); R.templateTests.runTests(); var F1000platform = new F1000.Platform({ name: "f1000research", displayName: "F1000Research", hostName: "f1000research.com", id: "1", editorialEmail: "[email protected]", infoEmail: "[email protected]", usePmcStats: true }); $(function(){R.ui.dropdowns('.dropdown-for-authors, .dropdown-for-about, .dropdown-for-myresearch');}); // $(function(){R.ui.dropdowns('.dropdown-for-referees');}); $(document).ready(function () { if ($(".cookie-warning").is(":visible")) { $(".sticky").css("margin-bottom", "35px"); $(".devices").addClass("devices-and-cookie-warning"); } $(".cookie-warning .close-button").click(function (e) { $(".devices").removeClass("devices-and-cookie-warning"); $(".sticky").css("margin-bottom", "0"); }); $("#tweeter-feed .tweet-message").each(function (i, message) { var self = $(message); self.html(linkify(self.html())); }); $(".partner").on("mouseenter mouseleave", function() { $(this).find(".gray-scale, .colour").toggleClass("is-hidden"); }); }); Sign In Remember me Forgotten your password? Sign In Cancel Email or password not correct. Please try again Please wait... $(function(){ // Note: All the setup needs to run against a name attribute and *not* the id due the clonish // nature of facebox... $("a[id=googleSignInButton]").click(function(event){ event.preventDefault(); $("input[id=oAuthSystem]").val("GOOGLE"); $("form[id=oAuthForm]").submit(); }); $("a[id=facebookSignInButton]").click(function(event){ event.preventDefault(); $("input[id=oAuthSystem]").val("FACEBOOK"); $("form[id=oAuthForm]").submit(); }); $("a[id=orcidSignInButton]").click(function(event){ event.preventDefault(); $("input[id=oAuthSystem]").val("ORCID"); $("form[id=oAuthForm]").submit(); }); }); If you've forgotten your password, please enter your email address below and we'll send you instructions on how to reset your password. The email address should be the one you originally registered with F1000. Email address not valid, please try again You registered with F1000 via Google, so we cannot reset your password. To sign in, please click here . If you still need help with your Google account password, please click here . You registered with F1000 via Facebook, so we cannot reset your password. To sign in, please click here . If you still need help with your Facebook account password, please click here . Code not correct, please try again Reset password Cancel Email us for further assistance. Server error, please try again. If your email address is registered with us, we will email you instructions to reset your password. If you think you should have received this email but it has not arrived, please check your spam filters and/or contact for further assistance. Please wait... Register $(document).ready(function () { signIn.createSignInAsRow($("#sign-in-form-gfb-popup")); $(".target-field").each(function () { var uris = $(this).val().split("/"); if (uris.pop() === "login") { $(this).val(uris.toString().replace(",","/")); } }); });

Text is read by the "Ask this paper" AI Q&A widget below. Extraction quality varies by source — PMC NXML preserves structure cleanly, OA-HTML may include some navigation residue, and OA-PDF can have broken hyphenation. The publisher copy (via DOI) is the canonical version.

My notes (saved in your browser only)

Ask this paper AI returns verbatim quotes from the full text · source: preprint-html

Answers must be backed by verbatim quotes from this paper's full text. Hallucinated quotes are dropped automatically; if no verbatim passage answers the question, we say so. How this works

Citation neighborhood (no data yet)

We don't have any in-corpus citations linked to this paper yet. This is a recent paper (2024) — citers typically take a year or two to land, and the OpenAlex reference graph may still be filling in.

Source provenance

europepmc
last seen: 2026-05-20T01:45:00.602351+00:00