Sample information curated by ChIP-Atlas

Antigen

Antigen Class
TFs and others
Antigen
Ctcf

Cell type

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

Attributes by original data submitter

Sample

source_name
Mouse embryonic stem cells v6.5
genotype
Wild Type
cell line
mESC v6.5
antibody
anti-CTCF; Active Motif 61311

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
mESCs were crosslinked in 1% formaldehyde for 2 minutes before quenching with 125 mM glycine. Crosslinked cells were lysed using Lysis Buffer 1 and Lysis Buffer 2 before resuspension in Sonication Buffer 1. Human chromatin (from HEK293T cells) was spiked in (to final 5%) prior to sonication for the indicated experiments. Sonication of nuclei was performed on a Covaris E220 with the following settings: Duty Factor 8, PIP/W 210, and 200 cycles per burst for 12 minutes. Chromatin fragments of 200-1,000 base pair size were generated. Antibody was incubated with 100uL Protein G Dynabeads for 6 hours. Unbound antibody was removed via washing, before incubation of antibody bound beads with chromatin overnight. Beads were then washed with Sonication Buffer 1 and Wash Buffers 1, 2, and 3. Chromatin was eluted and crosslinks were reversed overnight via incubation at 65C and addition of 5ul Proteinase K. Zymo ChIP DNA Clean and Concentrate kit (D5205) was used to purify DNA following Proteinase digestion. Sequencing libraries were prepared using NEBNext Ultra II DNA library prep kit for Illumina (E7645S).

Sequencing Platform

instrument_model
Illumina HiSeq 2500

mm10

Number of total reads
38659236
Reads aligned (%)
91.4
Duplicates removed (%)
7.2
Number of peaks
36118 (qval < 1E-05)

mm9

Number of total reads
38659236
Reads aligned (%)
91.3
Duplicates removed (%)
7.3
Number of peaks
36108 (qval < 1E-05)

Base call quality data from DBCLS SRA