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SRX739986: GSM1531258: oblong_polyA_RNA_Seq; Danio rerio; RNA-Seq
1 ILLUMINA (Illumina HiSeq 2000) run: 139.1M spots, 21.1G bases, 13Gb downloads

Submitted by: Gene Expression Omnibus (GEO)
Study: A high resolution sequencing and modeling identifies distinct dynamic RNA regulatory strategies
show Abstracthide Abstract
To monitor the relative regulatory contributions of RNA production, processing and degradation, we sampled RNA from mouse DCs every 15 minutes, for the first 3 hours of their response to LPS (13 samples in total), following a short (10 minute) metabolic labeling pulse with 4sU preceding the sampled time point. We isolated RNA from each of these samples in two ways. First, to comprehensively measure total RNA regardless of its transcription time, we isolated RNA depleted of rRNA. Second, to measure only newly made RNA, we isolated 4sU-labeled RNA, which could only have been transcribed during the 10 minute labeling pulse and is thus enriched for short-lived transcripts, including mRNA precursors and processing intermediates. We deeply sequenced each isolated RNA population to provide the necessary depth to study exons, introns, and splicing junctions across the transcriptome. We showed our approach general utility by also applying it to lincRNAs and early zebrafish development data Overall design: Time-course mRNA profiles of mouse DCs that were stimulated with LPS, and labeled with a short (10 minute) 4sU pulse (0-3 hours, 15 min. intervals, 13 time points).
Sample: oblong_polyA_RNA_Seq
SAMN03140395 • SRS727712 • All experiments • All runs
Organism: Danio rerio
Library:
Instrument: Illumina HiSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Mouse samples Total RNA was extracted with the miRNeasy kit's procedure (Qiagen). For preparation of RNA-Sequencing samples, we used 10 μg total RNA and removed rRNA with the RiboZero kit (Epicenter). After taking an aliquot of 100 ng ribosomal depleted total RNA, we followed with 4sU purification for the remainder of the sample as desceibed in Rabani et. al., Nat. Biotech., 2011. Zebrafish samples Total RNA was isolated using the standard Trizol (Invitrogen) protocol. Two rounds of PolyA+-RNA purification were performed for each sample, using the PolyA(Purist)TM-MAG kit (Ambion). The quality of the RNA and lack of contaminating ribosomal RNA were confirmed using the Agilent 2100 Bioanalyzer. We prepared the RNA-Seq libraries using the dUTP second strand (strand specific) protocol as described in Levin et. al, Nat. Methods, 2010.
Experiment attributes:
GEO Accession: GSM1531258
Links:
External link:
Runs: 1 run, 139.1M spots, 21.1G bases, 13Gb
Run# of Spots# of BasesSizePublished
SRR1621206139,089,93521.1G13Gb2014-12-16

ID:
1070824

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