# A protocol for chemical competence in phytopathogenic Ralstonia

## TL;DR

This paper provides a detailed protocol for making phytopathogenic Ralstonia bacteria chemically competent for transformation.

## Contribution

A novel and effective protocol for transforming Ralstonia solanacearum species complex strains using calcium chloride-induced chemical competence.

## Key findings

- The protocol successfully transformed four out of five tested Ralstonia solanacearum species complex strains.
- The method includes cold calcium chloride treatments and heat shock steps for efficient transformation.
- The protocol's efficacy is supported by quantitative results and logical development explanations.

## 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.