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Sample GSM1859525 Query DataSets for GSM1859525
Status Public on Jul 26, 2018
Title PCL2plus_Ter119neg_H3K27me3
Sample type SRA
 
Source name embryonic fetal liver
Organism Mus musculus
Characteristics strain background: C57Bl/6
genotype/variation: WT
developmental stage: E14.5
tissue source: embryonic fetal liver
sorted fraction: Ter119minus; CD71+Ter119-/lo
chip antibody: H3K27me3 Antibody
chip antibody vendor: Millipore
chip antibody cat.#: #07-449
chip antibody lot #: 2455635
Growth protocol Gene targeted mouse C57Bl/6 ES cells were obtained through EUCOMM and were aggregated with CD1 blastocysts to form chimeras.
Extracted molecule genomic DNA
Extraction protocol FL cells from e14.5 Pcl2+/+ and Pcl2-/- mice were isolated and CD71+Ter119-/lo and CD71+Ter119high fractions were sorted using a MoFlo sorter
For ChIP-seq, sorted cells were crosslinked with 1% formaldehyde for 10 minutes at room temperature. Samples were sheared using a Covaris sonicator until DNA reached a final size of 75-700bp. 10ug of antibody (anti-Pcl2, Genway; anti-H3K27me3, Millipore) was bound to pre-blocked Protein A magnetic beads (Millipore), combined with sonicated DNA and incubated overnight. After incubation, beads were collected and DNA-antibody complexes were eluted at 65oC. Crosslinks were reversed overnight at 65oC. Samples were treated with Proteinase K and RNase A and DNA was purified using phenol-chloroform. 500,000 cells per sample were used for both IP and control (IgG, SantaCruz).
 
Library strategy ChIP-Seq
Library source genomic
Library selection ChIP
Instrument model Illumina NextSeq 500
 
Data processing Bowtie v2.2.3 and MACS 1.3.7 were used for alignments and peak calling, respectively.
Genome_build: mm9
Supplementary_files_format_and_content: Bowtie generated alignments and MACS generated peak calling files for ChIPSeq data
 
Submission date Aug 24, 2015
Last update date May 15, 2019
Contact name William Stanford
E-mail(s) wstanford@ohri.ca
Phone 613-737-8899
Organization name Ottawa Hospital Research Institute
Lab Stanford
Street address 501 Smyth Road
City Ottawa
State/province Ontario
ZIP/Postal code K1H 8L6
Country Canada
 
Platform ID GPL19057
Series (2)
GSE72287 Mtf2-PRC2 control of canonical Wnt signaling is required for definitive erythropoiesis. [ChIP-seq]
GSE72288 Mtf2-PRC2 control of canonical Wnt signaling is required for definitive erythropoiesis
Relations
BioSample SAMN04009200
SRA SRX1162724

Supplementary data files not provided
SRA Run SelectorHelp
Raw data are available in SRA
Processed data are available on Series record

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