FULL INTERROGATION OF NUCLEASE DSBS AND SEQUENCING (FIND-SEQ)

摘要

Sensitive, unbiased methods for genome-wide detection of potential off-target nuclease cleavage sites in DNA, e.g., in cell type-specific genomic DNA samples.

主权项

1. A method of preparing a library of covalently closed DNA fragments, the method comprising:
providing DNA; optionally randomly shearing the DNA to a defined average length to provide a population of DNA fragments; preparing the fragments for end-ligation; ligating to the ends of the fragments a first hairpin adapter comprising at least a single deoxyuridine and a first primer site to prepare a population of ligated fragments; contacting the ligated fragments with a nuclease that induces double stranded breaks; and purifying the ligated fragments using an exonuclease, thereby preparing a library of covalently closed DNA fragments.