Fig. 3
Guide-it validation of the five sgRNAs synthesized for the generation of the GckrP446L point mutation. Cas9 protein is complexed with each sgRNA (B, D–G) and incubated with short double-stranded DNA fragments containing the protospacer target. Lanes A and C are controls and show the target template but no Cas9/sgRNA complex. The reactions are analyzed for cleavage by electrophoresis on agarose gel. L2 = 100 bp DNA molecular weight ladder (thick bands are 1000 and 500 bp). Protospacer sequences are detailed in Additional file 1: Table S1