GSM4847769: THLE-2_4-HNE; Homo sapiens; OTHER
After the irradiation or chemical treatment, genomic DNA was isolated using Quick-DNA Miniprep Plus Kit (Zymo Research; Irvine, CA) according to the manufacturer's instruction manual. Genomic DNAs were framgmeted to an averge length of 300 bp and circuliarized with T4 DNA ligase at 16°C overnight. Non-circularized linear DNA were removed with Plasmid-Safe ATP-Dependent DNAse treatment. CPD lesions and etheno-dA adducts were cleaved with combined treatment of DNA glycosylases, APE1, AP lyase and S1 nuclease to create double-strand breaks with ligatable ends. DNA was then A-tailed with Klenow fragment exo- (NEB) and then adaptor was ligated with NEBNext Ultra II Ligation Master Mix and followed by incubation with USER enzyme (NEB). The purified DNA libraries were amplified by PCR with NEBNext Ultra II Q5 Master mix and NEBNext i5 and i7 primers and used for DNA sequencing.