Sample information curated by ChIP-Atlas

Antigen

Antigen Class
Histone
Antigen
H3K4me3

Cell type

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

Attributes by original data submitter

Sample

source_name
1000 mESCs
development stage
cell strain
cell type
mESC
chip antibody
H3K4me3
genetic background
mixed background of MF1, 129/sv, and C57BL/6J strains

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
use microfludic chip to do ChIP The reagents for library preparation were provided by Illumina and were used according to the manufacturer’s protocol with appropriate modifications as described below. We purified DNA from the ChIP assay by AMPure XP beads and eluted the DNA by 20µl Resuspension Buffer. The DNA suspension was concentrated from 20µl to 5µl by evaporation. Then, 4µl End Repair Mix and 1µl Resuspension Buffer were added to the 5µl cleaned up DNA suspension before incubating at 30°C for 30 minutes and inactivating at 75°C for 15 minutes for end repairing. Adenylation of 3’ ends is performed by adding 8.4µl thawed A-Tailing Mix and 1.6µl Resuspension Buffer, incubating at 37°C for 30 minutes. We ligated adapters by adding 1.7µl of DNA ligase Mix, 3.1µl Resuspension Buffer and 0.2µl Adapter Index. 30°C for 10 minutes is the duration for adapter ligation. We added 1µl DNA which contains 100 ng of 5kb sized plasmid DNA as carrier, and then purified DNA twice by AMPure XP Beads.

Sequencing Platform

instrument_model
Illumina HiSeq 2000

mm10

Number of total reads
5080693
Reads aligned (%)
96.9
Duplicates removed (%)
4.4
Number of peaks
4060 (qval < 1E-05)

mm9

Number of total reads
5080693
Reads aligned (%)
96.8
Duplicates removed (%)
4.4
Number of peaks
4016 (qval < 1E-05)

Base call quality data from DBCLS SRA