Sample information curated by ChIP-Atlas

Antigen

Antigen Class
Histone
Antigen
H3K27me3

Cell type

Cell type Class
Cardiovascular
Cell type
Cardiac fibroblasts
NA
NA

Attributes by original data submitter

Sample

source_name
cFB
cell type
cardiac fibroblast
chip antibody
H3K27me3

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cells were resuspended with digestion buffer (50mM Tris-HCL, pH7.6, 1mM CaCl2, 0.2% triton X-100) and 5mM butyrate, 0.5mM PMSF, x proteinase inhibitor. The chromatins were digested with Mnase and enzyme was inactivated by adding the stop buffer (10mM Tris, pH7.6, 5mM EDTA). The chromatin was immunoprecipitated with antibodies specific to the H3K4me3 (ab8580, Abcam, Cambridge, UK), and H3K27me3 (07-449, Millipore, Billerica, MA) in RIPA buffer (10 mM Tris, pH7.4, 1mM EDTA, 0.1% SDS, 0.1% Na-Deoxycholate, and 1% Triton X-100). Bead complex was captured by magnetic stand and washed sequentially twice with RIPA buffer, RIPA + 0.3 M NaCl, and LiCl buffer (0.25 M LiCl, 0.5% NP40, 0.5% Na- Deoxycholate) and once with 1X TE + 0.2% Triton X-100, and 1X TE. Proteinase K digestion and phenol/chloroform extraction was performed to isolate the DNA. Sequencing libraries were manually generated for high-throughput sequencing. Either end of DNA strands was filled using the Epicentre DNA END-Repair kit (Epicentre Biotechnologies, Madison, WI). Adenine was added at 3’ ends by Taq DNA polymerase (New England Biolabs, Beverly, MA) under dATP conditions. Illumina’s adaptor oligomers was ligated at both ends using Quick Ligation kit (Qiagen, Germantown, MD). After purified twice, DNA libraries are amplified using Solexa primers (Illumina, San Diego, CA) and Phusion PCR master mix (Thermo Scientific, Hudson, NH).

Sequencing Platform

instrument_model
Illumina Genome Analyzer IIx

mm10

Number of total reads
9210089
Reads aligned (%)
90.1
Duplicates removed (%)
37.1
Number of peaks
142 (qval < 1E-05)

mm9

Number of total reads
9210089
Reads aligned (%)
90.1
Duplicates removed (%)
37.2
Number of peaks
139 (qval < 1E-05)

Base call quality data from DBCLS SRA