|
Status |
Public on Jun 02, 2012 |
Title |
Ezh2 RIP-deeper-seq biological replicate |
Sample type |
SRA |
|
|
Source name |
embryonic stem cells
|
Organism |
Mus musculus |
Characteristics |
strain: 129X (M.castaneusX129) cell type: embryonic stem cells rip antibody: anti-Ezh2 antibody
|
Growth protocol |
mouse embryonic stem cells were routinely grown and maintained in T25 or T75 flasks with LIF supplemented in the culture medium.
|
Extracted molecule |
total RNA |
Extraction protocol |
Cell nuclei were isolated, nuclear lysate was prepared and incubated with an anti-Ezh2 antibody. Afterwards, the RNA-protein complexes were immunoprecipitated with protein A agarose beads and the RNA fraction was extracted using Trizol reagent. Template switching method was used for the library construction per manufacturer's recommendation (Clontech's SMART cDNA Library Construction Manual).
|
|
|
Library strategy |
RIP-Seq |
Library source |
transcriptomic |
Library selection |
other |
Instrument model |
Illumina Genome Analyzer II |
|
|
Description |
2 lanes of sequence data from the biological replicate GSM489991
|
Data processing |
Image processing and base calling were performed using the Illumina RTA pipeline. Preprocessing: 3' adapter sequences were identified using cross_match and trimmed if a match of at least 5 bp was found. Reads matching mtDNA or rRNA sequences, reads < 12 bp long, reads containing N's, and homopolymer reads were removed. Alignment: reads were aligned to the mouse genome (mm9) using shortQueryLookup with a maximum of 1 error and no indels. Frequencies of unique reads were then tabulated. the 's.WT.BR.uniq.noribo.nohomop.tsv.gz' processed data file is derived from two Ezh2 RIP-deeper sequencing samples, and available on a Series records Genome_build: mm9 Supplementary_files_format_and_content: Frequencies of unique reads
|
|
|
Submission date |
Jun 01, 2012 |
Last update date |
May 15, 2019 |
Contact name |
Mark L Borowsky |
E-mail(s) |
borowsky@molbio.mgh.harvard.edu
|
Organization name |
MGH
|
Department |
Molecular Biology
|
Street address |
185 Cambridge Street
|
City |
Boston |
State/province |
MA |
ZIP/Postal code |
02114 |
Country |
USA |
|
|
Platform ID |
GPL9250 |
Series (1) |
GSE17064 |
Genome-wide identification of Polycomb-associated RNAs by RIP-seq |
|
Relations |
SRA |
SRX150919 |
BioSample |
SAMN01001654 |