Figure 5. Outline of genome editing procedure in B. subtilis .

(Adapted from Figure S2 [Burby and Simmons, 2017]). A. The first step is to transform B. subtilis with the editing plasmid. Inset shows wild-type mutS2 locus. B. After establishing isolates using colony purification, the isolates are cured of the editing plasmid. Inset depicts homology directed repair using the editing template on the plasmid. C. The final step is to verify loss of the editing plasmid and genotype isolates to verify the mutation was successfully introduced. Inset shows edited mutS2 locus.