alias: GSM4663375
title: GSM4663375: GM12878_4C_rep2; Homo sapiens; OTHER
library_construction_protocol: The cells were treated with 1% formaldehyde to crosslink, and the nuclei were isolated via resuspension in lysis buffer (500 μl 10 mM Tris-HCl pH 8.0, 10 mM NaCl, 0.2% Igepal CA-630 and 50 μl protease inhibitors). Nuclei were resuspended in 500 μl restriction enzyme buffer and permeabilized by incubation with 10% SDS (final concentration: 0.3%) for 37°C overnight. 500 units DpnII restriction enzyme were added and incubated overnight at 37 °C with shaking. Digestion was stopped after adding 1.6% SDS and incubated at 65 °C for 25 minutes. Samples were then diluted in 7.5 ml 1x ligation buffer and 100 Weiss unit of T4 DNA ligase was added. Samples were kept at 16 °C for 24 hours. The ligated chromatin was digested by proteinase K at 65 °C overnight. After purification, 16 parallel inverse PCR reactions, each containing 3.2 ug of circularized DNA, were performed.
identifiers.namespace: GEO
identifiers.id: GSM4663375