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
Embryonic stem cells
strain
129SvJae/C57BL/6J
material source
mESC cell line derived from F1 hybrid of 129SvJae/C57BL/6J
sorting
--
cell type
Embryonic stem cells
chip antibody
anti-H3K4me3 (Cell Signaling, catalog# 9751S, 11/2014, lot# 8)

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
mESCs (500 or 1000) or sorted HSC subtypes were mixed with ~5x108 DH5a E.coli and fixed in 1% formaldehyde for 10 min at room temperature. After fixation and washes, the mixtures were sonicated with a 1/16-inch probe for 15 min at 3 watts using a tip sonicator (Misonix sonicator 3000). The lysates were incubated with 2 microliter H3K27me3 antibody (Millipore 07-449) or 1 microliter H3K4me3 antibody (Cell Signaling 9751S) overnight at 4oC. To block any non-specific chromatin absorption, 60 microliter protein G magnetic beads (Life Technologies 10004D) or 10 microliter M-280 streptavidin beads (Life Technologies 11206D) were pre-incubated with ~5x108 fixed and sonicated E.coli lysate or the same E.coli lysate plus 5 ng carrier biotin-DNA, respectively, at 4oC overnight. These treated beads were combined and used to ChIP H3K27me3 or H3K4me3 in the sonicated E.coli+mESC or E.coli+HSC lysates for 4 h at 4oC. The beads were then recovered using a magnetic stand. All washing, de-crosslinking, and library building steps were performed following Illumina True-Seq protocol with 12 PCR amplification cycles. After end repair, A-tailing, and adapter ligation steps, 0.25 micro-M blocker oligo was added during final library amplification.

Sequencing Platform

instrument_model
Illumina HiSeq 2000

mm10

Number of total reads
61141284
Reads aligned (%)
17.6
Duplicates removed (%)
38.3
Number of peaks
23664 (qval < 1E-05)

mm9

Number of total reads
61141284
Reads aligned (%)
17.6
Duplicates removed (%)
38.5
Number of peaks
23585 (qval < 1E-05)

Base call quality data from DBCLS SRA