2024-09-21
ENA first public
ERC000011
ENA-CHECKLIST
University of Zaragoza
INSDC center name
public
INSDC status
ERS20599309
SRA accession
E-MTAB-14255:Sample-1
Submitter Id
ArrayExpress
broker name
not collected
collection date
Protocols: Bacterial growth from three blood agar plates of H. pylori 26695 incubated for 48 h at 37 °C under microaerobic conditions was aseptically resuspended in brain heart infusion broth supplemented with 4% foetal bovine serum (BHI + FBS) and adjusted to 107 colony forming units per millilitre (CFU/mL). BHI broth cultures were incubated microaerobically at 37 °C for 24 h. Next, aliquots of 12 mL were exposed in triplicate to 4 × MIC of the HsrA inhibitor and further incubated in the same above conditions until log10 CFU/mL resulted diminished in one unit. At this time, bacterial growth was stopped by adding 1.5 mL ice-cold RNA stop solution (95% absolute ethanol, 5% acid-buffered phenol), and subsequently incubated on ice. Control samples were exposed to the same amount of dimethyl sulfoxide (DMSO, vehicle) instead inhibitor. Helicobacter cells from all samples were harvested by centrifugation (4,000 rpm at 4 ºC for 10 min), pellets were quickly frozen by immersion in liquid nitrogen and stored at -80 ºC.  Frozen cell pellets from two biological replicates of each treatment condition were properly submitted to GENEWIZ (Azenta Life Sciences, Leipzig, Germany) in dry ice.  RNA-seq analysis was performed by GENEWIZ as follow. RNA-seq libraries were prepared according to standard protocols via rRNA depletion, RNA fragmentation and random priming, reverse transcription, 5´-phosphorylation and dA-tailing of cDNAs, adaptor ligation and PCR enrichment.
description
not collected
geographic location (country and/or sea)
not applicable
isolate
Helicobacter pylori 26695
organism
Helicobacter pylori 26695
organism
whole organism
organism part
1
replicate
Helicobacter pylori 26695
scientific_name
ATCC 700392 (aka 26695)
strain
Sample-1
title