Sample information curated by ChIP-Atlas

Antigen

Antigen Class
TFs and others
Antigen
Rnf2

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, tamoxifen treated, RING1B ChIP
chip antibody
RING1B ab (Atsuta et al., 2001; PMID 11289226)
cell line
MS12 KDM2Bfl/fl ES cells
passage
p10-p20

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
To prepare material for ChIP-Seq, immunoprecipitation was performed overnight at 4°C with approximately 3 µg of antibody and chromatin corresponding to 5 × 106 cells. Antibody bound chromatin was isolated on protein A magnetic Dynabeads (Invitrogen, Carlsbad, CA) and washes were performed with low salt buffer (0.1% SDS, 1% Triton, 2 mM EDTA, 20 mM Tris-HCl (pH 8.1), 150 mM NaCl), high salt buffer (0.1% SDS, 1% Triton, 2 mM EDTA, 20 mM Tris-HCl (pH 8.1), 500 mM NaCl), LiCl buffer (0.25 M LiCl, 1% NP40, 1% Deoxycholate, 1 mM EDTA, 10 mM Tris-HCl (pH 8.1)) and TE buffer (x2) (10 mM Tris-HCl (pH 8.0), 1 mM EDTA). To prepare ChIP-seq material, ChIP DNA was eluted, and cross-links reversed at 65oC, then samples were then sequentially treated with RNase and proteinase K before being purified on a PureLink PCR micro column (Invitrogen). ChIP sequencing libraries were generated as described previously (Blackledge et al., 2010) and sequenced on the Illumina HiSeq2000 platform with 51 bp reads. ChIP sequencing experiments were performed in biological triplicates for KDM2B and RING1B and biological duplicates for SUZ12. For mRNA-seq analysis 1 μg of purified total RNA was used to first isolate polyA plus RNA and then a directional library was prepared using the NEBNext UltraTM Directional RNA library Prep Kit for Illumina (NEB, Ipswich, MA). Performed by the High-Throughput Sequencing centre at the Wellcome Trust Centre for Human Genetics (Oxford, United Kingdom) according to standard Illumina library generation protocol.

Sequencing Platform

instrument_model
Illumina HiSeq 2000

mm10

Number of total reads
29219921
Reads aligned (%)
97.4
Duplicates removed (%)
17.1
Number of peaks
6971 (qval < 1E-05)

mm9

Number of total reads
29219921
Reads aligned (%)
97.0
Duplicates removed (%)
17.1
Number of peaks
6962 (qval < 1E-05)

Base call quality data from DBCLS SRA