Sample information curated by ChIP-Atlas

Antigen

Antigen Class
Histone
Antigen
H3K27ac

Cell type

Cell type Class
Embryonic fibroblast
Cell type
NIH/3T3
Primary Tissue
Embryo
Tissue Diagnosis
Normal

Attributes by original data submitter

Sample

source_name
NIH3T3 cell culture
strain background
NA
tissue/chip rxn
1x10^6 cells/ChIP
age
NA
chip antibody
H3K27ac (abcam ab4729)
genotype or treatment
48 hours LIF + 400 nm purmorphamine

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cells were fixed for 15 minutes at room temperature with 1% formaldehyde then lysed. After cell lysis, chromatin was sheared in shearing buffer containing 0.25% SDS using a Covaris S2 focused ultrasonicator. Sheared chromatin was incubated with antibody coated dynabead preparations (1ug antibody with 20ul beads) overnight. Beads were washed 5 times with RIPA buffer (1% NP40, 0.7% Sodium Deoxycholate, 1mM EDTA pH8, 50mM Hepes-KOH pH7.5, 2% w/v Lithium Chloride) and then eluted at 70°C for 15 minutes in elution buffer (50mM TRIS pH 8, 10mM EDTA pH 8, 1% SDS). Crosslinks were reversed overnight at 70°C and chromatin was purified and concentrated by phenol chloroform extraction and EtOH precipitation. One third of each ChIP sample was used for library preparation. Library preparation was done according to the Broad Pond method, (PMCID: PMC3091298) using the NEBNext library preparation kit (NEB E6040L) using non-standard 15 cycles of PCR amplification.

Sequencing Platform

instrument_model
Illumina HiSeq 4000

mm10

Number of total reads
35109068
Reads aligned (%)
83.7
Duplicates removed (%)
13.2
Number of peaks
34841 (qval < 1E-05)

mm9

Number of total reads
35109068
Reads aligned (%)
83.6
Duplicates removed (%)
13.3
Number of peaks
34860 (qval < 1E-05)

Base call quality data from DBCLS SRA