GeneTailor™ Site-Directed Mutagenesis System - FAQs

After a PCR mutagenesis protocol, how is the plasmid repaired after it is transformed into E. coli?

The detailed mechanism is still not very clear, but involves the E. coli double strand break repair system. The key steps are:

(1) An exonuclease, like RecBCD, removes nucleotides from the ends to generate a single-stranded region.

(2) Strand invasion with homologous regions occurs.

(3) Repair enzymes including nucleases, ligase, and DNA polymerases repair gaps and/or nicks.