Abstract
Here, we present a protocol for the transformation of Ralstonia solanacearum species complex (RSSC) strains by calcium chloride-induced chemical competence. We include a step-by-step protocol with media and solution recipes, quantitative results of the protocol's efficacy for four of five tested RSSC strains, and an explanation of the logic underlying the protocol development. The protocol involves sequentially treating overnight cultures with a cold calcium chloride solution and a cold calcium chloride/magnesium chloride/glycerol solution, transforming through alternating heat shocks and ice incubations, and recovering cells via an outgrowth in rich medium.
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Abstract
Here, we present a protocol for the transformation of Ralstonia solanacearum species complex (RSSC) strains by calcium chloride-induced chemical competence. We include a step-by-step protocol with media and solution recipes, quantitative results of the protocol's efficacy for four of five tested RSSC strains, and an explanation of the logic underlying the protocol development. The protocol involves sequentially treating overnight cultures with a cold calcium chloride solution and a cold calcium chloride/magnesium chloride/glycerol solution, transforming through alternating heat shocks and ice incubations, and recovering cells via an outgrowth in rich medium.
- Received:
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Funding
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National Science Foundation
(Award 2336557)
- Principal Award Recipient: Tiffany M. Lowe-Power
-
National Institute of Food and Agriculture
(Award 2024-67013-43303)
- Principal Award Recipient: Tiffany M. Lowe-Power
-
U.S. Department of Agriculture
(Award 1023861)
- Principal Award Recipient: Tiffany M. Lowe-Power
-
National Institute of Food and Agriculture
(Award 2024-67011-42914)
- Principal Award Recipient: Matthew L. Cope-Arguello
-
University of California, Davis
- Principal Award Recipient: Neha N. Prasad
-
University of California, Davis
- Principal Award Recipient: Tabitha C. Cowell
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