Sample information curated by ChIP-Atlas

Antigen

Antigen Class
Histone
Antigen
H3K4me3

Cell type

Cell type Class
Pluripotent stem cell
Cell type
EpiSC
NA
NA

Attributes by original data submitter

Sample

source_name
Epiblast stem cells
cell type
E5.75 derived EpiSCs
passages
20-25
strain
OCT4-deltaPE-GFP x C57BL/6
chip antibody
H3K4me3 (Abcam 8580, lot# GR95375-2)

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cells were fixed with 1% formaldehyde at 37C for 10min, quenched with glycine at 37C for 5 min. Fixed cells were sonicated using Epishear (Active Motif) to achieve 200-700 size chromatin fragments. Solubilized chromatin was immunoprecipiated with antibody against H3K4me3 (Abcam 8580) and H3K27me3 (Millipore 07-449). Antibody-chromatin complexes were pulled down using Dynabeads protein A (Invitrogen), washed and then eluted. After cross-linking reversal, RNase and proteinase K treatment, immunoprecipiated DNA was purified using AMPure beads (Beckman Coulter). ChIP DNA were end-repaired and 5' phosphorylated using T4 DNA Polymerase, Klenow and T4 Polynucleotide Kinase (Enzymatics). A single Adenine was added to 3' ends by Klenow (3-->5' exo-), and double-stranded Bioo Illumina Adapters (Bioo Scientific) were ligated to the ends of the ChIP fragments. Adapter-ligated ChIP DNA fragments were subjected to 15 cycles of PCR amplification using Q5 polymerase (NEB). AMPure beads were used to purify DNA after each step (Beckman Coulter). Pooled libraries were sequenced on the NextSeq 500 for single-end 75bp using high-output flowcell or on the HiSeq 2000 for single-end 36bp according to manufacturer's instructions.

Sequencing Platform

instrument_model
NextSeq 500

mm10

Number of total reads
16162199
Reads aligned (%)
90.9
Duplicates removed (%)
19.3
Number of peaks
19157 (qval < 1E-05)

mm9

Number of total reads
16162199
Reads aligned (%)
90.7
Duplicates removed (%)
19.4
Number of peaks
19142 (qval < 1E-05)

Base call quality data from DBCLS SRA