|
| Status |
Public on Mar 13, 2014 |
| Title |
Y1F-S7A 1 |
| Sample type |
SRA |
| |
|
| Source name |
S. pombe cells
|
| Organism |
Schizosaccharomyces pombe |
| Characteristics |
strain: rpb1CTD-Y1F-S7A
genetic markers: h- ade6 ura4-D18 leu1–32
|
| Growth protocol |
Two biological replicates of the WT, Y1F, S2A, T4A, S7A,Y1F-S7A, S2A-S7A and T4A-S7A cultures were grown in YES (yeast extracts plus supplements) media at 30°C till they reached OD600 0.4 -0.7.
|
| Extracted molecule |
polyA RNA |
| Extraction protocol |
Cells were harvested and RNA was isolated using the Qiagen RNeasy kit. RNA quality was assessed on a Bioanalyzer instrument (Agilent) and 1µg of total RNA was used to prepare polyA+ RNA using the Illumina TruSeq RNA sample kit .
RNA-seq libraries were prepared from polyA+ RNA using the Illumina TruSeq stranded total RNA Sample Prep Kit according to the manufacturers protocol. Indexed libraries were normalized and pooled (8 samples per lane) for paired-end sequencing performed using an Illumina HiSeq 2000 instrument at the Weill Cornell Medical College Genome Core Facility (in New York).
|
| |
|
| Library strategy |
RNA-Seq |
| Library source |
transcriptomic |
| Library selection |
cDNA |
| Instrument model |
Illumina HiSeq 2000 |
| |
|
| Description |
Y1F-S7A_1 (Name in expression table)
|
| Data processing |
Paired-end reads of length 51 bases (each part) originating from each sample were aligned using Bowtie 0.12.7 to the S. pombe genome sequence (Ensembl S. pombe, Build EF1) as well as to its corresponding exon-exon junctions database (only thesecond part of the paired-end reads was considered). Up to 3 mismatches were allowed. Reads that matched multiple loci were removed from further analysis and the resultant alignment files were processed to generate ‘pile-ups’ against each chromosome.
Searches were performed against the genome sequence combined with a dataset of known exon-exon junctions as defined by Ensembl S. pombe release-13. To ensure that a 51-base read mapped to a splice junction, only the last 45 bases of the first exon and the first 45 bases of the second exon were considered (if the exon length exceeded 45 nucleotides). In this way, reads that overlapped a junction by less than 6 nucleotides were excluded. Reads that matched to more than one junction or elsewhere in the genome were also discarded.
Genome_build: Ensembl S. pombe, Build EF1, version 58.1a
Supplementary_files_format_and_content: Expression table and RPKM summary for all samples tab-delimited
|
| |
|
| Submission date |
Nov 14, 2013 |
| Last update date |
May 15, 2019 |
| Contact name |
Danny Asher Bitton |
| E-mail(s) |
d.bitton@ucl.ac.uk
|
| Phone |
+44(0)203-108-1604
|
| Organization name |
University College London
|
| Department |
Department of Genetics, Evolution & Environment
|
| Lab |
Genome Regulation/Bähler Lab
|
| Street address |
Darwin Building, Gower Street
|
| City |
London |
| ZIP/Postal code |
WC1 6BT |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL13988 |
| Series (1) |
| GSE52370 |
Individual letters of the RNA polymerase II CTD code govern distinct gene expression programs in fission yeast |
|
| Relations |
| BioSample |
SAMN02402418 |
| SRA |
SRX377472 |
