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SRX14201136: GSM5903001: t0_control; Pseudomonas aeruginosa; OTHER
1 OXFORD_NANOPORE (PromethION) run: 4.9M spots, 2.1G bases, 1.7Gb downloads

External Id: GSM5903001_r1
Submitted by: Laboratory of Gene Technology, Biosystems, KU Leuven
Study: Development of ONT-cappable-seq to unravel the transcriptional landscape of Pseudomonas phages
show Abstracthide Abstract
We developed ONT-cappable-seq, a specialized long-read RNA sequencing technique that allows end-to-end sequencing of primary prokaryotic transcripts using the Nanopore sequencing platform. We applied ONT-cappable-seq to study the transcriptional landscape of Pseudomonas aeruginosa phage LUZ7, leading to a comprehensive genome-wide map of viral transcription start sites, terminators and complex operon structures that fine-regulate gene expression. At the same time, it provides new insights in the RNA biology of LUZ7 and paves the way for more in depth transcription studies that can help unveil the complex layers of phage-host interactions. Overall design: Full-length transcriptional profiling of P. aeruginosa US449 infected with phage LUZ7 at different time points (0, 5, 10 and 20 minutes) using ONT-cappable-seq.
Sample: t0_control
SAMN25995410 • SRS12021715 • All experiments • All runs
Library:
Name: GSM5903001
Instrument: PromethION
Strategy: OTHER
Source: TRANSCRIPTOMIC
Selection: other
Layout: SINGLE
Construction protocol: Total RNA was extracted using hot-phenol extraction, followed by DNAse treatment. The enriched samples were subjected to an enrichment procedure based on streptavidin beads to enrich for primary transcripts. For the control samples the enrichment steps were omitted. Total RNA was enriched for primary transcripts using an adapted version of the (SMRT)-cappable-seq enrichment protocol. In parallel, control samples were not enriched but subjected to similar environmental conditions. The RNA was reverse transcribed, PCR amplified and barcoded according to Oxford Nanopore Technology cDNA-PCR protocol (SQK-PCS109 combined with SQK-PBK004). The amplified cDNA samples were pooled together in a final library and loaded on a promethION flowcell.
Runs: 1 run, 4.9M spots, 2.1G bases, 1.7Gb
Run# of Spots# of BasesSizePublished
SRR180488724,855,9872.1G1.7Gb2022-06-24

ID:
20064554

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