DNA sequencing of Bacteroides phage BV01: culture supernatant from B. vulgatus ATCC 8482
Phage DNA was prepared from overnight TYG culture supernatants collected after centrifugation and filtration with a 0.22 m filter and concentrated by centrifugation with 30,000 MWCO Corning Spin-X UF 20 Concentrators (Corning, NY) or by tangential flow filtration with a Vivaflow 50R 30,000 MWCO Hydrosart filter (Sartorius, Gottingen, Germany). Supernatants were treated with 200 g/mL DNase I and 1 g/mL RNase A for 1 hr at room temperature to remove unprotected DNA and RNA. Virions were disrupted with 1% SDS and 1 mg/mL proteinase K for 2 hr at 55C. DNA was further isolated using the same phenol-chloroform-isoamyl alcohol extraction and ethanol precipitation procedures as for cellular DNA. DNA libraries were constructed with the Nextera XT Library Preparation Kit and Index Kit (Illumina, San Diego, CA). DNA libraries were pooled and sequenced on an Illumina HiSeq2500 instrument. Fastq files were generated from demultiplexed reads with bcl2fastq Conversion Software (Illumina, San Diego, CA). Reads were trimmed using the A5ud pipeline.
SUB7892085
D1