|
| Status |
Public on Jul 26, 2018 |
| Title |
PCL2plus_Ter119plus_PCL2 |
| 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: Ter119plus; CD71+Ter119high
chip antibody: Pcl2 (also known as Mtf2/M96) antibody
chip antibody vendor: GenwayGWB
chip antibody cat.#: FA7207
chip antibody lot #: QC3873-39693
|
| 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 |
SAMN04009191 |
| SRA |
SRX1162729 |
