GSM5261773: XR-Seq A2780; Homo sapiens; OTHER
cells were harvested, collected by centrifugation and resuspended in lysis buffer and lysed using a Dounce homogenizer. XR-seq was performed as previously described (Hu. et al Genes and Dev. 2016, Adar S. et al PNAS 2016). Briefly, primary excision products were pulled-down by TFIIH co-immunoprecipitation using anti-p62 antibody (Santa-Cruz Biotechnology, sc-293) and anti-p89 antibody (Santa-Cruz Biotechnology, sc-292). Adapters were ligated on both ends of the excision oligomer and immunoprecipitated using cisplatin-specific antibody (abcam, 103261). Cisplatin-induced damages were reversed in-vitro by incubating the samples with 200 mM NaCN at 65°C overnight. DNA was PCR amplified and purified by native polyacrylamide gel electrophoresis (PAGE) to create the library.
GEO
GSM5261773