Supplementary Figure 3: Optimization of in vitro circularization conditions with uracil-containing stem loop adapters and a PCR amplicon.
From: CIRCLE-seq: a highly sensitive in vitro screen for genome-wide CRISPR–Cas9 nuclease off-targets
Qiaxcel capillary electrophoretic traces of intramolecular ligation, exonuclease treatment, and restriction enzyme digestion. The observed electrophoretic mobility shift is consistent with circularization. An exonuclease-resistant population of circular DNA molecules is observed after Plasmid-Safe treatment. Digestion with BamHI restriction enzyme linearizes the circularized DNA and results in the expected shift in mobility.
