Figure 4. Linearization of plasmid DNA by target site cleavage with rSaCas9
Plasmid DNA containing cloned target sequences (pJET_CCD8) was incubated with purified rSaCas9 and one of ten different sgRNAs (T1-T10) for site-directed DNA cleavage. Fifteen-microliter aliquots of the cleavage reactions were loaded on an agarose gel, and nucleic acids were visualized by ethidium bromide staining. Samples without enzyme (w/o rSaCas9) or without guide RNA (w/o sgRNA) served as controls. The closed arrowhead indicates the slower-migrating linearized plasmid, and the open arrowhead indicates the undigested plasmid DNA.
Figure 1