Sample information curated by ChIP-Atlas

Antigen

Antigen Class
Histone
Antigen
H3K27ac

Cell type

Cell type Class
Pluripotent stem cell
Cell type
ES cells
NA
NA

Attributes by original data submitter

Sample

source_name
Embryonic stem cell (ESC)
cell type
ESC
line
C57BL/6
genotype
TKO+Dnmt3a1
antibody
H3K27ac (Active Motif 39133)

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
mESCs were pre-plated to deplete MEF feeders before collection. Cells were fixed with 1% formaldehyde after which chromatin was isolated and sonicated to 150-300 bp using Bioruptor (Diagenode). Sonicated chromatin was precleared with Protein A/G Dynabeads then incubated in antibody overnight. Chromatin was then immunoprecipitated with Protein A/G Dynabeads for > 2 hours. ChIP was washed successively by two rounds of low-salt buffer, two rounds of high-salt buffer, two rounds of LiCl buffer, and finally two rounds of TE buffer. ChIP DNA was eluted in TE, reverse crosslinked overnight, treated with RNase A, then proteinase K, and finally extracted with phenol-chloroform and ethanol precipitation. ChIP-seq libraries were prepared using KAPA Hyper Prep Kit per manufacturer's recommendations. Libraries were quantified using Qubit Fluorometer and Bioanalyzer High Sensitivity DNA Analysis Kit individually. Libraries were pooled and sequenced using the Illumina HiSeq 2000 machine as either 50-bp or 100-bp single-end sequencing reads.

Sequencing Platform

instrument_model
Illumina HiSeq 2000

mm10

Number of total reads
22387627
Reads aligned (%)
69.4
Duplicates removed (%)
19.5
Number of peaks
13830 (qval < 1E-05)

mm9

Number of total reads
22387627
Reads aligned (%)
69.2
Duplicates removed (%)
19.7
Number of peaks
13534 (qval < 1E-05)

Base call quality data from DBCLS SRA