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SRX481483: GSM1340089: DNAPKCS METHANOL HS ChipSeq technical replicate 1 and 2; Homo sapiens; ChIP-Seq
2 ILLUMINA (Illumina HiSeq 2500) runs: 74.7M spots, 3.8G bases, 2.3Gb downloads

Submitted by: Gene Expression Omnibus (GEO)
Study: Role of Polycomb group proteins in the DNA damage response – a reassessment
show Abstracthide Abstract
We have investigated whether components of Polycomb repressive complex 1 (PRC1) are recruited to double-strand breaks (DSBs) generated by inducible expression of the AsiSI restriction enzyme in normal human fibroblasts. Using chromatin immunoprecipitation and deep sequencing (ChIP-seq), which detects PRC1 proteins at common sites throughout the genome, we did not find evidence for recruitment of PRC1 components to AsiSI-induced DSBs. In contrast, the S2056 phosphorylated form of DNA-PKcs and other DNA repair proteins were detected at a subset of AsiSI sites that are predominantly at the 5’ ends of transcriptionally active genes. Our data question the idea that PcG protein recruitment provides a link between DSB repairs and transcriptional repression. Overall design: Single chromatin preparations from treated and untreated cells (+/- OHT), immunoprecipitated with two antibodies (pDNA-PKcs and MEL18) were sequenced as technical replicates, along with the corresponding input DNAs
Sample: DNAPKCS METHANOL HS ChipSeq technical replicate 1 and 2
SAMN02673414 • SRS566221 • All experiments • All runs
Organism: Homo sapiens
Library:
Instrument: Illumina HiSeq 2500
Strategy: ChIP-Seq
Source: GENOMIC
Selection: ChIP
Layout: SINGLE
Construction protocol: DNA samples were end repaired, poly-A tailed and Illumina single end adapters were ligated following the standard Illumina protocol with minor adjustments. Agencourt AMPure XP beads at 0.8x ratio were used to size select out adapter dimers after adapter ligation. The Illumina kit Phusion enzyme was replaced by Kapa HiFi HotStart ready mix. Post PCR, AMPure XP beads were used at a 1:1 ratio to maintain size integrity and to allow use of the Invitrogen SizeSelect E-gel system. Samples were finally purified with QAIquick gel extraction kit and quality controlled on the DNA 1000 BioAnalyser 2100 chip before clustering and subsequent 50-51bp single end sequencing on the Illumina HiSeq 2500.
Experiment attributes:
GEO Accession: GSM1340089
Links:
External link:
Runs: 2 runs, 74.7M spots, 3.8G bases, 2.3Gb
Run# of Spots# of BasesSizePublished
SRR118393521,071,5801.1G612.8Mb2014-07-06
SRR118393653,614,0102.7G1.7Gb2014-07-06

ID:
671556

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