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SRX474180: GSM1331424: LB 2.0 B2 TEX pos L2 HS2; Escherichia coli str. K-12 substr. MG1655; RNA-Seq
1 ILLUMINA (Illumina HiSeq 2000) run: 6.6M spots, 616.7M bases, 382.6Mb downloads

Submitted by: Gene Expression Omnibus (GEO)
Study: Identification and validation of antisense RNAs in Escherichia coli
show Abstracthide Abstract
In this study transcriptional start sites (TSS) of E. coli were determined for several growth conditions Overall design: To detect the complement of transcripts expressed from E. coli, we collected two independent biological replicates (B1 and B2 samples) from MG1655 wild type strain grown to exponential (OD 600 ~0.4) or stationary phase (OD 600 ~2.0) in LB medium (samples LB 0.4 and LB 2.0, respectively) as well as grown to exponential phase (OD 600 ~0.4) in M63 minimal glucose medium (sample M63 0.4). For all six samples, total RNA was extracted and subjected to differential RNA-seq (dRNA-seq) library preparation for primary transcriptome analysis as described previously (Sharma et al., 2010). Specifically, prior to cDNA library construction half of each RNA sample was treated with 5' terminator exonuclease (+TEX samples), which degrades RNAs containing a 5'-mono-phosphate (5'-P) and, thus, enriching enriches for primary transcripts containing 5'-tri-phosphates (5'-PPP). The other half of each sample was left untreated (-TEX samples) and thus contains both primary transcripts (5'-PPP) and processed RNAs (5'-P).
Sample: LB 2.0 B2 TEX pos L2 HS2
SAMN02646737 • SRS560169 • All experiments • All runs
Library:
Instrument: Illumina HiSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Hot phenol (Sharma et al. 2010, Bloomberg 1990, EMBO J) The libraries were generated by Vertis Biotechnologie AG (Munich, Germany). The samples were poly(A)-tailed by using poly(A) polymerase. The 5'-PPP were removed using tobacco acid pyrophosphatase (TAP) followed by the ligation of the RNA adapter to the 5'-monophosphate of the RNA. First-strand cDNA synthesis was performed with an oligo(dT)-adapter primer and the M-MLV reverse transcriptase. The resulting cDNA was PCR-amplified to reach a concentration of 20-30 ng/μl using a high fidelity DNA polymerase. The cDNA was purified using the Agencourt AMPure XP kit (Beckman Coulter Genomics) and was analyzed by capillary electrophoresis. The primers used for PCR amplification were designed for TruSeq sequencing according to the instructions of Illumina
Experiment attributes:
GEO Accession: GSM1331424
Links:
External link:
Runs: 1 run, 6.6M spots, 616.7M bases, 382.6Mb
Run# of Spots# of BasesSizePublished
SRR11739806,561,093616.7M382.6Mb2014-10-20

ID:
654670

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