alias: GSM2047013
title: GSM2047013: A375, rep2; Homo sapiens; OTHER
library_construction_protocol: 10 million cells were fixed with 1% formaldehyde and nuclei pellets were isolated after cell lysis with cold lysis buffer (50mM Tris-Hcl pH 7.5, 150mM NaCl, 5mM EDTA, 0.5% NP-40, 1% TritonX-100) supplemented with protease inhibitors. First step digestion was performed over-night at 37°C with 200unit HindIII enzyme. After phenol-chloroform extraction, DNA was ligated over-night at 16°C by T4 DNA ligase. Following de-crosslinking, DNA was processed for second digestion with 50units of DpnII enzyme for over-night at 37°C. After final ligation 4C template DNAs were quantified by Qubit dsDNA High sensitivity kit 4C library was prepared as described previously (Splinter et al. 2012). Tert promoter specific primers with Illimuna Nextera adapters were used for libraries.
identifiers.namespace: GEO
identifiers.id: GSM2047013