Sample information curated by ChIP-Atlas

Antigen

Antigen Class
TFs and others
Antigen
Dnmt3a

Cell type

Cell type Class
Embryonic fibroblast
Cell type
MEF
Tissue
Embryonic Fibroblast
Lineage
primaryCells
Description
Mouse Embryonic Fibroblast

Attributes by original data submitter

Sample

source_name
MEF
chip antibody
Dnmt3a (Abcam ab13888, mouse)
strain
C57BL/6
genotype
Dnmt3a(+/-)

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
ChIP-seq was performed as described (Deshpande et al 2014 Cancer Cell). 1-2×106 MEF cells were crosslinked using 1% formaldehyde, washed in 1 mL of wash buffer (20 mM Tris pH8.0, 167mM NaCl, 5mM EDTA, 1% SDS) and lysed in lysis buffer (20 mM Tris pH8.0, 167mM NaCl, 5mM EDTA, 0.66% SDS, 1% Triton X-100). Chromatin was sheared to 200-400 bp using Covaris E220 sonicator and used for chromatin immunoprecipitation (ChIP) with Dnmt3a-specific antibodies ab13888 mouse mAbs and ab2850 rabbit pAbs (Abcam) and Protein-A (Life Technologies) magnetic beads. After washing, the immune complexes were eluted and DNA fragments recovered after de-crosslinking at 65°C for 4 hours and used for TruSeq libraries generated using NEBNext kit (New England BioLabs). Libraries were sequenced using HiSeq2500 (Illumina) to generate approximately 4×107 single end (SE)50 reads per sample.

Sequencing Platform

instrument_model
Illumina HiSeq 2000

mm10

Number of total reads
22727774
Reads aligned (%)
95.6
Duplicates removed (%)
16.4
Number of peaks
385 (qval < 1E-05)

mm9

Number of total reads
22727774
Reads aligned (%)
95.3
Duplicates removed (%)
16.5
Number of peaks
426 (qval < 1E-05)

Base call quality data from DBCLS SRA