Sample information curated by ChIP-Atlas

Antigen

Antigen Class
Histone
Antigen
H2A.XS139ph

Cell type

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

Attributes by original data submitter

Sample

source_name
gH2AX_HG-mESCs_ChIP-seq
protocol
25 mM glucose
strain
129S4/SvJae
cell type
Undifferentiated embryonic stem cells
chip-antibody
α-γH2AX (Rabbit polyclonal antibody, Abcam, ab2893)

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
ChIP-enriched DNA was obtaind by using ChIP-IT Express Enzymatic (Active Motif, 53009) and S40Gc specific antibody (20B2). Total RNA was isolated from cells using Direct-zol RNA Kit (Zymo Research, R2052) according to the manufacturer’s instructions. The ends of 50 ng of ChIP-enriched DNA and Input DNA were repaired using T4 DNA polymerase (New England Biolabs, M0203) and phosphorylated with T4 polynucleotide kinase (New England Biolabs, M0201). A single adenine base was added to the 3 prime-end with Klenow fragments (New England Biolabs, M2012). TruSeq DNA adapters (Illumina, FC-121-2001/-2002) were ligated to the fragments with T4 DNA ligase (New England Biolabs, M2200). Ligation products between 200 and 600 bp were purified on AMpure XP beads (Beckman Coulter, A63880) to remove unligated adapters and subjected to 14 cycles of PCR amplification by KAPA Library Amplification Kit (KAPA BIOSYSTEMS, KK2611). RNA-seq libraries were constructed using TruSeq Strand mRNA LT Sample Prep Kit (Illumina) according to the manufacturer’s instructions. Completed libraries were quantified using an Agilent 2000 BioAnalyzer system.

Sequencing Platform

instrument_model
Illumina HiSeq 2000

mm10

Number of total reads
19394311
Reads aligned (%)
78.4
Duplicates removed (%)
12.7
Number of peaks
176 (qval < 1E-05)

mm9

Number of total reads
19394311
Reads aligned (%)
78.2
Duplicates removed (%)
12.8
Number of peaks
137 (qval < 1E-05)

Base call quality data from DBCLS SRA