GSM3223782: Input BAF60cKO CM R1; Mus musculus; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Input control
Antigen
Input control
Cell type
Cell type Class
Pluripotent stem cell
Cell type
mESC derived cardiac cells
NA
NA
Attributes by original data submitter
Sample
source_name
cardiomyoctes
cell type
ESC-derived cardiac myocytes
genotype
BAF60c KO
passage
15-20
strain
W4/129S6
chip antibody
input
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
ChIP was performed following O'Geen et al., 2011, Wamstad et al., 2012 with modifications. Briefly, 50 million cells were crosslinked with disuccinimidyl gulatarate (45 min) followed by formaldehyde (15min) and quenched, chromatin was digested with MNase to mono and di-nucleosomes, BRG1 was immuniprecipitated using anti-BRG1 antibody (ab110641, lot no. )and washed extensively and DNA eluted followed by reverese crosslinking, proteinase and RNAse treatment and ampure purification of DNA (Serandour et al., 2013). Libraries were prepared mannually. Briefly, DNA was end-repaired using the End-IP repair kit (Epicenter, # ER81050). DNA were A-tailed with Klenow fragment (3' to5' exo minus) and dATP. Barcoded Illumina adapters were ligated and DNA was PCR amplified with Illumina primers for 14 cycles. 300- 500bp library fragments were size selected in an agarose gel and purified. Libraries were quantified by Qubit and analyzed in a Bioanalyser. Libraries were diluted and sequenced in a NEBNextSeq sequencing system following manufacturer's instructutions.