NgAgo and mutants were incubated with plasmid DNA and 5'P guide sequences targeting the plasmid. Nicked/cleaved products were amplified via primer extension, which terminates at the nick site, before
We report the PAMs of NmeCas9 using a cell-free TXTL-based cleavage assay. By adding randomized PAM library and NmeCas9-gRNA in vitro, functional PAM sequences were cleaved, while non-functional PAMs
