A Rapid Bacteriophage DNA Extraction Method

Bacteriophages (phages) are intensely investigated as non-antibiotic alternatives to circumvent antibiotic resistance development as well as last resort therapeutic options against antibiotic resistant bacteria. As part of gaining a better understanding of phages and to determine if phages harbor pu...

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Bibliographic Details
Published inMethods and protocols Vol. 1; no. 3; p. 27
Main Authors Jakočiūnė, Džiuginta, Moodley, Arshnee
Format Journal Article
LanguageEnglish
Published Switzerland MDPI 27.07.2018
Subjects
Protocol
genome
DNA extraction
spin column
fast
phage
sequencing
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Summary:Bacteriophages (phages) are intensely investigated as non-antibiotic alternatives to circumvent antibiotic resistance development as well as last resort therapeutic options against antibiotic resistant bacteria. As part of gaining a better understanding of phages and to determine if phages harbor putative virulence factors, whole genome sequencing is used, for which good quality phage DNA is needed. Traditional phage DNA extraction methods are tedious and time consuming, requiring specialized equipment e.g., an ultra-centrifuge. Here, we describe a quick and simple method (under four hours) to extract DNA from double stranded DNA (dsDNA) phages at titers above 1.0 × 1010 plaque-forming units (PFU)/mL. This DNA was suitable for library preparation using the Nextera XT kit and sequencing on the Illumina MiSeq platform.
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ISSN:2409-9279
2409-9279
DOI:10.3390/mps1030027