biostudies-literatureYang YNatural Science Foundation of China838698https://www.ebi.ac.uk/biostudies/studies/S-EPMC8989280biostudies-literatureUnknown13<i>Escherichia coli recA</i> ^- strains are usually used for cloning to prevent insert instability <i>via</i> RecA-dependent recombination. Here, we report that <i>E. coli</i> BW25113 (<i>recA</i> ⁺) competent cells prepared by using a previously reported transformation and storage solution (TSS) had 100-fold or higher transformation efficiency than the commonly used <i>E. coli</i> cloning strains, including XL1-Blue MRF'. The cloning success rates with <i>E. coli</i> BW25113 were 440 to 1,267-fold higher than those with <i>E. coli</i> XL1-Blue MRF' when several inserts were assembled into four vectors by using a simple DNA assembly method. The difference was in part due to RecA, as the <i>recA</i> deletion in <i>E. coli</i> BW25113 reduced the transformation efficiencFrontiers in microbiology<i>Escherichia coli</i> BW25113 Competent Cells Prepared Using a Simple Chemical Method Have Unmatched Transformation and Cloning Efficiencies.PMC8989280318700853196113301591951202Wang MLiu HXia YYang YXun LZhao RYu Qfalse<i>Escherichia coli</i> BW25113 Competent Cells Prepared Using a Simple Chemical Method Have Unmatched Transformation and Cloning Efficiencies.<i>Escherichia coli recA</i> ^- strains are usually used for cloning to prevent insert instability <i>via</i> RecA-dependent recombination. Here, we report that <i>E. coli</i> BW25113 (<i>recA</i> ⁺) competent cells prepared by using a previously reported transformation and storage solution (TSS) had 100-fold or higher transformation efficiency than the commonly used <i>E. coli</i> cloning strains, including XL1-Blue MRF'. The cloning success rates with <i>E. coli</i> BW25113 were 440 to 1,267-fold higher than those with <i>E. coli</i> XL1-Blue MRF' when several inserts were assembled into four vectors by using a simple DNA assembly method. The difference was in part due to RecA, as the <i>recA</i> deletion in <i>E. coli</i> BW25113 reduced the transformation efficienc2022-01-01T00:00:00Z20222025-04-05T00:27:38.935Z2025-04-05T00:27:38.935ZS-EPMC89892803540148410.3389/fmicb.2022.838698