|
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 |