GSM2974249: WTZT16KIDNEY rep2; Mus musculus; OTHER
Tissue were lysed gently and small molecular weight DNA, consisting mostly of excised 26-27 mers was separated from chromosomal high molecular weight DNA by centrifugation Briefly, the excised oligomers are immunopricipitated with anti-TFIIH (anti-XPB and anti-p62) antibodies because the excision product is released in a relatively stable complex with TFIIH, which also protects the excision product from degradation by non-specific nucleases. This is followed by extraction of the excised oligomers and ligation to adaptors and a second immunoprecipitation with either anti-CPD or anti-BPDE antibodies. Human DNA polymerases η and κ were used for primer extension in the presence of purified excised oligonucleotides containing either CPD or BPDE-dG damages, as polymerases η and κ can bypass CPD and BPDE-dG damage respectively in an error-free manner. This is followed by PCR amplification with index primers and NGS sequencing on HiSeq 2500 platform.
GEO
GSM2974249