Sample information curated by ChIP-Atlas

Antigen

Antigen Class
Input control
Antigen
Input control

Cell type

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

Attributes by original data submitter

Sample

source_name
C59_input
cell line
JM8.N4 mouse embryonic stem cell line
genotype
FLAG-Halo-mCTCF/mRad21-SNAPf-V5 Knock-in
ctcf protein
wild type
chip antibody
none

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cells were washed twice with ice-cold PBS, scraped, centrifuged for 10 min at 4000 rpm, resuspended in cell lysis buffer (5 mM PIPES, pH 8.0, 85 mM KCl, and 0.5% NP-40, 1 ml/15 cm plate) and incubated for 10 min on ice. Lysates were then centrifuged for 10 min at 4000 rpm. Nuclear pellets were resuspended in 6 volumes of sonication buffer (50 mM Tris-HCl, pH 8.1, 10 mM EDTA, 0.1% SDS), incubated on ice for 10 min, and sonicated to obtain DNA fragments below 2000 bp in length (Covaris S220 sonicator, 20% Duty factor, 200 cycles/burst, 100 peak incident power, 50 cycles of 30" on and 30" off). Sonicated lysates were cleared by centrifugation and 400-800 μg of chromatin was diluted in RIPA buffer (10 mM Tris-HCl, pH 8.0, 1 mM EDTA, 0.5 mM EGTA, 1% Triton X-100, 0.1% SDS, 0.1% Na-deoxycholate, 140 mM NaCl) to a final concentration of 0.8 μg/μl, precleared with Protein A sepharose (GE Healthcare) for 2 hours at 4°C and immunoprecipitated overnight with 4-8 μg of antibodies. About 15% of the precleared chromatin was saved as input. Immunoprecipitated DNA was purified with the Qiagen QIAquick PCR Purification Kit, eluted in 60 μl of water and used for library preparation. Solexa rapid prep (see linked manuscript)

Sequencing Platform

instrument_model
Illumina HiSeq 4000

mm10

Number of total reads
79044865
Reads aligned (%)
98.4
Duplicates removed (%)
25.1
Number of peaks
535 (qval < 1E-05)

mm9

Number of total reads
79044865
Reads aligned (%)
98.1
Duplicates removed (%)
25.1
Number of peaks
594 (qval < 1E-05)

Base call quality data from DBCLS SRA