alias: GSM3272773
title: GSM3272773: XR-seq_Liver_1; Mus musculus; OTHER
library_construction_protocol:  Genomic DNA were extracted using PurLink Genomic DNA kit (Thermo).  Four hours after injecting cisplatin or saline solution (control group), the mice were sacrificed by carbon dioxide exposure, the kidneys, liver, lung and spleen were removed and washed extensively with cold phosphate buffered saline (PBS), and then homogenized them.  The chromatin fraction was then pelleted.  The supernatants (containing low molecular DNA-protein fragments) were harvested and immunoprecipitated (IP) with anti-TFIIH (p89 antibody (G-10), and p62 antibody (H-10), Santa-Cruz Biotechnology).  The eluted DNA were isolated using phenol-chloroform and ethanol precipitation. The low-molecular excision products were immunoprecipitated with anti-TFIIH (p89 antibody and p62 antibody), and ligated to adaptors on the both ends.  After second round immunoprecipitated with cisplatin-specific anti-body, the Pt-DNA adducts were reversed by incubating in NaCN (200 mM) over-night.  Then the damage free DNAs were amplified by PCR to get XR-seq libraries and sequenced using an Illumina HiSeq 4000 with 50-bp single-end read.
identifiers.namespace: GEO
identifiers.id: GSM3272773
attributes.tag: GEO Accession
attributes.value: GSM3272773