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Sample GSM1032193 Query DataSets for GSM1032193
Status Public on Dec 04, 2012
Title H3K27ac ChIP: KAP1 KD TI
Sample type SRA
Source name H3K27ac ChIP: KAP1 KD TI
Organism Mus musculus
Characteristics strain: C57BL/6
cell type: Embryonic stem cells (ES3)
time-point: 4 days post puromycin selection of KAP1 shRNA vector
chip antibody: None (Total Input)
Growth protocol ES cells were cultured in Glasgow Minimum Essential Medium (GMEM, Sigma: G5154) with sodium pyruvate (used at 1mM, Sigma: S8636), MEM non-essential amino acids (used at 1x, Gibco: 11140-035), L-glutamine (used at 2mM, Gibco: 25030-024), 2-mercaptoethanol (used at 0.1mM, Sigma), ES cell tested FBS (Gibco: 16141-079) and leukaemia inhibitory factor (LIF, used at 1,000 units/ml, Chemicon: ESG1107). Cells were grown on 0.2% gelatin (Sigma: 48723-500G-F)-coated plates and split every two days.
Extracted molecule genomic DNA
Extraction protocol ES cell samples were washed 2x (in PBS + 2% FCS), counted to normalize by cell number, cross-linked (ten minutes rotation in 1% formaldehyde), quenched with glycine (at 125mM on ice), washed 3x (PBS) and pelleted at 10e7 cells per ependorf. Pellets were lysed, resuspended in 1ml sonication buffer on ice (10mM Tris pH 8, 200mM NaCl, 1mM EDTA, 0.5mM EGTA, 0.1% NaDOC, 0.25% NLS and protease inhibitors), transferred to glass 12x24mm tubes (Covaris: 520056) and sonicated (Covaris settings: 20% duty cycle, intensity 5, 200cyles/ burst, 30 minutes). Sonication was then assessed by reverse cross-linking overnight in the presence of proteinase K and RNase, followed by DNA extraction and quantification on a Bioanalyzer (Agilent 2100 machine). Samples were also checked for the absence of single-stranded DNA by Exonuclease I treatment.
Library strategy ChIP-Seq
Library source genomic
Library selection ChIP
Instrument model Illumina HiSeq 2000
Data processing Raw reads were mapped to the mouse genome and transcriptome using Bowtie
Peaks were called using MACS
For histone modifications, enriched regions were defined using the ChIP-part suite
Genome_build: mm9
Submission date Nov 06, 2012
Last update date May 15, 2019
Contact name Adamandia Kapopoulou
Phone 0041216930940
Organization name EPFL
Department Sciences de la Vie
Lab Trono
Street address Station 15
City Lausanne
ZIP/Postal code 1015
Country Switzerland
Platform ID GPL13112
Series (1)
GSE41903 TRIM28 repression of retrotransposon-based enhancers is necessary to preserve transcriptional dynamics in embryonic stem cells
SRA SRX202992
BioSample SAMN01801732

Supplementary data files not provided
SRA Run SelectorHelp
Raw data are available in SRA
Processed data not provided for this record

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