Described in Garcia-Tuñon I et al. Stem Cell Res.(2011)7(1), 1-16. and in Pérez-Palacios et al., 2016.
strain
Mak et al., 2012
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
ChIPseq protocol from Pérez-Palacios et al., 2016. 5 x 106 cells for each experimental condition were chemically crosslinked for 15 min at room temperature by the addition of fresh 37% formaldehyde solution to a final concentration of 1% (v/v). After the addition of glycine to a final concentration of 125 mM to stop the crosslink, cells were rinsed twice with PBS and harvested by scraping. Cells were resuspended in lysis buffer (50mM Tris-HCl (pH 8.1), 10mM EDTA, 1% (w/v) SDS) and protease inhibitors (CompleteTM, Roche) and sonicated in a Diagenode sonicator (30 second pulses, 30 second pause between pulses). Libraries were prepared according to Illumina's instructions. Libraries were sequenced on the Genome Analyzer following the manufacturer's protocols using a TruSeq (TM) SBSv5 sequencing kit.. ChIP-Seq, instrument version Illumina Hi-Seq 2000, sequencing cycles 50