Fig. 1
From: Rapid characterization of CRISPR-Cas9 protospacer adjacent motif sequence elements
Schematic for identification of PAM preferences by Cas9 cleavage in vitro. a Initial plasmid library with randomized PAM (green box) is cleaved with Cas9 complex and 3′ dA overhangs are added. b Adapters with 3′ dT overhang (blue box) are ligated to both ends of the cleavage product. c Primers are utilized to enrich for PAM-sided cleaved products by PCR. d After PCR enrichment, DNA fragments are purified and Illumina compatible anchors and barcodes are ‘tailed-on’ through two rounds of PCR (gray boxes) and Illumina deep sequenced