Sample information curated by ChIP-Atlas

Antigen

Antigen Class
TFs and others
Antigen
Brd2

Cell type

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

Attributes by original data submitter

Sample

source_name
K3R3 Brd2 ChIP-seq
strain background
V6.5 (129SvJaexC57BL/6)
cell type
Embryonic stem cells (ESCs)
transfected with
H2A.Z(K3R3)-YFP
genotype/variation
expressing a ubiquitin mutant version of H2A.Z
chip antibody
Brd2 (Bethyl A302-583A, lot 1)

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
The ChIP protocol has been adapted from previous studies (Lee et al., 2006; Boyer et al., 2006, Creyghton et al., 2008) with the following modification. Diagenode Bioruptor was used to sonicate with 30 cycles of 30 sec on, 30 sec off. The samples were sonicated in 15ml polystyrene tubes at 4°C while samples were immersed in ice cold water. ~200 ng of DNA was submitted to SPRI-works Fragment Library System I (Beckman Coulter) for each library prepared. Briefly, the DNA is subjected to size selection (200-400bp) with magnetic beads, end repaired, then a single adenine nucleotide is added to allow for directional ligation of adaptors. For this study, a 1:100 dilution of Single End read adapters (Illumina) was used in the ligation reaction. Each sample was then amplified for 19 cycles, using the Illumina protocol, adding additional bases from the PCR primers that aid sequences to anneal to the Illumina Genome Analyzer flow cell. Specific primers were used for 3’ barcoding, adding 6 nt barcodes to uniquely identify each sample (V6.5 Brd2 Rep 1: TACCGC, Rep 2: GTCCAG, WT Brd2 Rep 1: TTCAGA, WT Brd2 Rep 2: TGCTGT, WT Brd4 Rep 1: ACGGTG, WT Brd4 Rep 2: CATCAC, K3R3 Brd2 Rep 1: GTAACA, K3R3 Brd2 Rep 2: GGATCT, K3R3 Brd4 Rep 1: TCCGGG, K3R3 Brd4 Rep 2: AAGCTC). The samples were then purified on a Qiagen MinElute column, and libraries were quantified by Quant-it DNA Assay (Invitrogen, Q-33120), and examined for proper size and structure by Bioanalyzer (Agilent) and qPCR

Sequencing Platform

instrument_model
Illumina HiSeq 2000

mm10

Number of total reads
19240486
Reads aligned (%)
98.1
Duplicates removed (%)
69.6
Number of peaks
8589 (qval < 1E-05)

mm9

Number of total reads
19240486
Reads aligned (%)
97.9
Duplicates removed (%)
69.8
Number of peaks
8547 (qval < 1E-05)

Base call quality data from DBCLS SRA