Sample information curated by ChIP-Atlas

Antigen

Antigen Class
Unclassified
Antigen
Unclassified

Cell type

Cell type Class
Pluripotent stem cell
Cell type
P19
Tissue
Embryo
Disease
Teratocarcinoma; Embryonal Carcinoma

Attributes by original data submitter

Sample

source_name
RA-treated P19 EC cells
cell type
EC-derived neural progenitor cells
cell line
P19

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cells were scraped in phosphate buffered saline 48 hours after RA addition and were pelleted at 100 g before genomic DNA extraction using a DNeasy Blood and Tissue kit (Qiagen, France). Twenty micrograms of DNA in 300 ml of TE buffer (Tris 10 mM, EDTA 0.1 mM, pH 8.0) were sonicated with a Bioruptor (Diagenode, Belgium) to yield 200 to 500 bp fragments. Each glucosylation and biotinylation reaction was run using reagents from the Hydroxymethyl Collector kit (ref. 55013, Active Motif, Belgium) and 500 ng of sonicated DNA. For SCL-seq, 16 reactions were captured on streptavidin-coated magnetic beads (ref. 11205D, Invitrogen). After 5 washes and elution from the beads according to the manufacturer's protocol, the captured DNA was purified, precipitated and pooled for sequencing library preparation using the TruSeq ChIP Sample Prep Kit (Illumina, ref. IP-202-1012). For SCL-exo, after 6 washes in RIPA buffer (50 mM HEPES pH 7.6; 1 mM EDTA; 0.7% Na-Deoxycholate; 1% NP-40; 0.5 M LiCl) and 2 washes in Tris 10 mM pH 8, the DNA-beads complex was processed as previously described [19]: end polishing, ligation of the P7 exo-adapter, nick repair, lambda and RecJf exonuclease digestion, elution, P7 primer extension, ligation of the P5 exo-adapter, PCR amplification, and finally gel-size selection. The exonuclease-digested DNA was eluted from the beads by incubation in 100 ml of Elution Buffer (95% formamide, 10 mM EDTA) at 90 ⁰C for 5 min, followed by DNA precipitation and resuspension in 20 ml of water. The SCL-seq and SCL-exo libraries were quantified using the KAPA library quantification kit for Illumina sequencing platforms (KAPA Biosystems, KK4824) and 40 bp single-end sequenced in a single lane of a HiSeq 2000 (Illumina) following the manufacturer's protocol.

Sequencing Platform

instrument_model
Illumina HiSeq 2000

mm10

Number of total reads
22554206
Reads aligned (%)
97.3
Duplicates removed (%)
7.8
Number of peaks
482 (qval < 1E-05)

mm9

Number of total reads
22554206
Reads aligned (%)
97.1
Duplicates removed (%)
7.8
Number of peaks
512 (qval < 1E-05)

Base call quality data from DBCLS SRA