Figure 3. CATCH cleavage and cloning of long genomic sequences of various lengths from different genomic locations.

(a) A total of five sgRNA pairs were designed to target segments of different lengths (50, 75, 100, 150 and 200 kb, respectively) in the E. coli DH5a str. WPN25-sfGFP genome, all containing an sfGFP gene. The genome location, target sequences, PAM sequences and cleavage sites are detailed and highlighted. (b) E. coli chromosomes in agarose gel plugs were digested by Cas9 with the corresponding sgRNA pairs and analysed by PFGE. M, marker.