alias: 853
title: DNA sequencing of Bacteroides vulgatus mutant: prophage BV01 deletion strain
design: Cells were pelleted from 5 mL overnight culture in TYG by centrifugation at 4,000  g for 5 min at 4C, resuspended in 0.5 mL TE buffer (10 mM Tris, 1 mM EDTA), and lysed by adding sodium dodecyl sulfate (SDS) and proteinase K (GoldBio, Olivette, MO) to final concentrations of 0.07% and 300 g/mL, respectively, and incubating for 2 hr at 55C. Cellular material was removed by washing twice in an equal volume of buffered phenol, phenol-chloroform-isoamyl alcohol (VWR, Radnor, PA), and DNA precipitated with 100% ethanol in the presence of 0.3M sodium acetate at -20C overnight. DNA pellets were washed with 70% ethanol, dried, and resuspended in TE buffer. DNA libraries were constructed with the Nextera XT Library Preparation Kit and Index Kit (Illumina, San Diego, CA) or the NEBNext Ultra II FS kit for Illumina (New England Biolabs, Ipswich, MA). DNA libraries were pooled and sequenced on an Illumina HiSeq5000, HiSeq2500 and MiSeq instruments. Fastq files were generated from demultiplexed reads with bcl2fastq Conversion Software (Illumina, San Diego, CA). Reads were trimmed and assembled using the A5ud pipeline (Riley et al., 2017).
identifiers.namespace: SUB7896863
identifiers.id: 853