Damage-seq of hela-s3 for (6-4)PP, late phase, replicate A
Genomic DNA were fragmented by sonication, followed by end-repair, dA-tailing and adaptor ligation by NEBNext UltraII DNA library preparation kit. Adaptor-ligated fragments were danatured and subjected to IP with damage specific antibodies. Primer extension was performed to detect the exact position of damage on these purified fragments, because DNA polymerase stalls when encountering a lesion. Read-through products were removed by subtractive hybridization, then the rest blocked products were attached to the second adaptor. The ligation products were PCR-amplified with index primers and sequenced by Illumina Hiseq system.
SUB6971447
Hela_64_DS_NA_late_12_A