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Sample GSM549565 Query DataSets for GSM549565
Status Public on Mar 02, 2012
Title H3K9me2 WT 2
Sample type SRA
 
Source name MEF
Organism Mus musculus
Characteristics strain: C57BL/6
cell type: Fibroblasts from day 13.5 embryos immortalized with SV40-LTA
genotype: G9a f/f, ERT2-Cre
passages: Passaged 12-15 times
chip antibody: H3K9me2
vendor: Abcam (ab1220)
lot: 625300
Treatment protocol Primary MEFs were immortalized at passage 2. At passage 5, MEFs were treated witheither EtOH as a vehicle control or Tamoxifen to induce deletion of G9a.
Growth protocol MEFs were grown in 10 cm plates with DMEM medium containing 15% FBS, Pen/Strep, Glutamine, NEAA and 2-mercaptoethanol. After immortalization with SV40-LTA, MEFs were grown in 15 cm plates with the same medium except FBS was 10%.
Extracted molecule genomic DNA
Extraction protocol Lysates were clarified from sonicated nuclei and histone-DNA complexes were isolated with antibody. Libraries were prepared according to Illumina's instructions accompanying the DNA Sample Kit (Part# 0801-0303). Briefly, DNA was end-repaired using a combination of T4 DNA polymerase, E. coli DNA Pol I large fragment (Klenow polymerase) and T4 polynucleotide kinase. The blunt, phosphorylated ends were treated with Klenow fragment (3' to 5' exo minus) and dATP to yield a protruding 3- 'A' base for ligation of Illumina's adapters which have a single 'T' base overhang at the 3’ end. After adapter ligation DNA was PCR amplified with Illumina primers for 15 cycles and library fragments of ~250 bp (insert plus adaptor and PCR primer sequences) were band isolated from an agarose gel. The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on the Genome Analyzer following the manufacturer's protocols.
 
Library strategy ChIP-Seq
Library source genomic
Library selection ChIP
Instrument model Illumina Genome Analyzer II
 
Description chromatin IP against H3K9me2 in G9a f/f cells
Data processing Raw data processed using onboard SCS/RTA 2.6, alignments performed using Bowtie keeping only unique reads with 2 or fewer mismatches in 36bp. Alignments summed in 100bp windows for wig files and integrated profiles.
 
Submission date Jun 02, 2010
Last update date May 15, 2019
Contact name Terry Fang
E-mail(s) tfang@rockefeller.edu
Phone 212-327-8265
Fax 212-327-8258
Organization name Rockefeller University
Street address 1230 York Ave
City New York
State/province NY
ZIP/Postal code 10065
Country USA
 
Platform ID GPL9250
Series (1)
GSE22102 Histone H3 lysine 9 di-methylation as an epigenetic signature of the interferon response (sequencing)
Relations
SRA SRX021898
BioSample SAMN00014669

Supplementary file Size Download File type/resource
GSM549565_Terry_1_WT_redo_fastq_prefilter_bowtie.sam.bed.wig.gz 23.4 Mb (ftp)(http) WIG
SRA Run SelectorHelp
Processed data provided as supplementary file
Raw data are available in SRA

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