Sample information curated by ChIP-Atlas

Antigen

Antigen Class
Input control
Antigen
Input control

Cell type

Cell type Class
Others
Cell type
Fibroblasts
NA
NA

Attributes by original data submitter

Sample

source_name
Fibroblast
strain
C3H/An
tissue
Subcutaneous connecive tissue
cell type
A9 fibroblast
chip antibody
-

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
For ChIP assays, lysates were clarified from sonicated nuclei (Diagenode), and DNA-protein complexes were immunoprecipitated using antibodies conjugated to Dynabeads (ThermoScientific). For Viral Chromosome Conformation Capture assays, MVM infected cells were crosslinked in 2 percent formaldehyde for 10 minutes, quenched in 0.125 M glycine for 5 minutes, collected and lysed in NP-40 containing 3C Lysis buffer for 10 minutes before being subjected to primary restriction enzyme digestion overnight with 400 U of HindIII. Following digestion, the restriction enzyme was inactivated and samples were subjected to intramolecular ligation with T4 DNA Ligase at room temperature. Samples were then reverse crosslinked and purified, before digesting in the secondary restriction enzyme, NlaIII. NlaIII was heat inactivated, samples were purified by phenol:chloroform purification, and circularized using T4 DNA Ligase. Samples were purified using a PCR Purification Kit (Qiagen), and equivalent amounts of DNA was used to perform inverse PCR with primers complementary to the MVMp genome. Inverse PCR samples were diluted 1:100 in TE, before being used as templates for nested-inverse PCR reactions. 6 independent neste-inverse PCR reactions were pooled, PCR purified using a Qiagen kit and submitted for library preparation and high-throughput sequencing. Libraries were prepared using the NEBNext Ultra II DNA Library Prep Kit for Illumina (cat # E7645L) according to manufacturer's instructions. Twelve samples were pooled per run and samples were analyzed by 75 bp single-end sequencing on the NextSeq 500 (Illumina).

Sequencing Platform

instrument_model
NextSeq 500

mm10

Number of total reads
28077883
Reads aligned (%)
93.6
Duplicates removed (%)
12.9
Number of peaks
208 (qval < 1E-05)

mm9

Number of total reads
28077883
Reads aligned (%)
93.4
Duplicates removed (%)
12.9
Number of peaks
225 (qval < 1E-05)

Base call quality data from DBCLS SRA