GSM5550778: HCT116 cells, siCtrl_pAMNase_2; Homo sapiens; OTHER
Mnase activity was induced by addition of 2mM CaCl2 for 30 min on ice. Chromatin fragments were released by incubating the samples for 20 minutes at 37°C and centrifugation at 16.000×g for 10 minutes at 4°C. Supernatants were incubated at 70°C in the presence of 0.1% SDS and 5 µg proteinase K. DNA was cleaned-up using phenol/chloroform extraction NGS library preparation was performed using NuGEN´s Ovation Ultralow System V2 (M01379 v5). Libraries were prepared with a starting amount of 1 ng of DNA and were amplified in 12 PCR cycles. Libraries were profiled in a High Sensitivity DNA on a 2100 Bioanalyzer (Agilent technologies) and quantified using the Qubit dsDNA HS Assay Kit, in a Qubit 2.0 Fluorometer (Life technologies).