GSM9696216: DMSO_noTDP1_Biotin; Homo sapiens; Human gammaherpesvirus 8; OTHER
Genomic DNA from 5 × 107 cells treated with DMSO or PNR07-02 was purified and treated with 5 μg of pancreatic RNase (Roche) followed by with or without 1 μg of TDP1 in 80 mM KCl, 50 mM Tris–HCl pH 8.0, 2 mM EDTA,2 mM DTT for 1 h at 37 °C, adjusted to 0.5% SDS and incubated for 1 h at 55 °C with proteinase K (200 μg/mL). DNA was extracted twice with phenol and precipitated with ethanol in the presence of 2 M ammonium acetate. The Epicentre DNA END-Repair kit (Epicentre Biotechnologies, Madison, WI, USA) was used to generate blunt-ended DNA. SSB and DSB DNA libraries were prepared using NEBNext utra II DNA library prep kit for Illumina (NEB). The purified DNA was used directly for cluster generation and sequencing analysis using the Illumina Genome Analyzer (Illumina, San Diego, USA) following manufacturer protocols.