Systematic Regional Bias is Widespread in ChIP-seq

ABSTRACT Robust and reproducible results are essential for confident scientific analysis. We demonstrate that transcription factor (TF) Chromatin Immunoprecipitation coupled with sequencing (ChIP-seq) suffers from systematic bias that may threaten its reproducibility: 80% of 200+ condition-matched, dual-replicate experiments in ENCODE contain genomic regions of systematic bias. We observe this regional bias even between replicates produced within the same experiment, resulting in thousands of unreplicated peaks, which often contain valuable biological data. We provide evidence that regional bias may lead to qualitative differences in TF biology inferred by different experiments; we discovered eight TFs with binding activity in compact chromatin that was identified by one experiment, yet systematically absent from others. To mitigate the effects of bias, we derive simple but effective metrics to quantify the quality of data within biased regions and demonstrate that they can be used for the robust integration of data from multiple experiments. Competing Interest Statement The authors have declared no competing interest.