title: GSM4151334: ChIP_EoV5_A_input; Mus musculus; ChIP-Seq
library_construction_protocol: Day 4 EomesV5/V5 (CCE) and WT (CCE) D4 EBs were dissociated using Trypsin-LE (Gibco) and ~3-4x107 cells were cross-linked for 10 min at RT with 1% (v/v) formaldehyde, followed by quenching with 125mM glycine. Nuclei were recovered and lysed to obtain chromatin, which was then sonicated to 200–500 bp, pre-cleared with protein G Dynabeads (Thermofisher Scientific) and ~175 ug of chromatin was immunoprecipitated with 10μg of anti-V5 (Abcam cat# ab9116, lot# GR: 322448-4) bound to protein G Dynabeads overnight on a rotator at 4ºC. Dynabeads were washed and immune complexes were eluted in IP elution buffer (1% SDS, 0.1M NaHCO3), reverse crosslinked overnight at 65ºC, RNaseA treated for 1.5hrs at 42ºC, proteinase K treated for 2hrs at 45ºC and DNA was recovered using a Zymo column kit. DNA was multiplexed and paired end sequencing was performed on a single lane of an Illumina HiSeq4000 platform.
attributes.tag: GEO Accession
attributes.value: GSM4151334