ERC000011
ENA-CHECKLIST
2022-08-18T12:14:33Z
ENA-FIRST-PUBLIC
2022-08-18T12:14:33Z
ENA-LAST-UPDATE
SAMEA110666181
External Id
Ghent University Department of Pharmaceutical Sciences Laboratory of Pharmaceutical Microbiology
INSDC center name
2022-08-18T12:14:33Z
INSDC first public
2022-08-18T12:14:33Z
INSDC last update
public
INSDC status
ERS12763761
SRA accession
E-MTAB-12091:Sample 16
Submitter Id
ArrayExpress
broker name
Protocols: Evolved bacterial lineages were grown in liquid BHI medium over night, pelleted, and immediately submitted to DNA extraction. Genomic DNA was extracted from 2 mL planktonic cultures. The pellet was resuspended in TE buffer (10 mM Tris-HCl pH 8 + 1 mM EDTA pH 8). 100 µL cell suspension was added to 500 µL lysis buffer (50 mM Tris-HCl pH 8, 70 mM EDTA pH 8, 1% sodium dodecyl sulfate) with 0.5 µg/mL pronase (Roche, Germany) containing acid-washed glass beads (Sigma-Aldrich, USA). After vortexing, the tubes were incubated at 37°C for 1 hour. 200 µL saturated ammonium acetate was added, vortexed, then 600 µL chloroform was added, and vortexed. After centrifuging for 5 minutes 400 µL of the top aqueous phase was transferred to a new tube containing 1 mL 100% ethanol. This was mixed by inversion and centrifuged for 5 minutes. The pellet was washed with 70% ethanol, and air-dried. The DNA was dissolved in low-EDTA-TE buffer (10 mM Tris-HCl pH 8 + 0.1 mM EDTA pH 8 + 0.5 µg/mL) containing RNase A (Qiagen, Germany), incubated at 37°C for 1 hour. NEBNext Ultra II library prep with 460-1000 ng input, according to the protocol with size selection using ampure XP beads after adapter ligation. PCR-free.
description
wild type genotype
genotype
biofilm liquid BHI medium from beads
growth condition
Pseudomonas aeruginosa
organism
Pseudomonas aeruginosa
organism
whole organism
organism part
Pseudomonas aeruginosa
scientific_name
AA2
strain
Sample 16
title