A Rapid Bacteriophage DNA Extraction Method

Methods and Protocols

Moodley, Arshnee¹  JakočiÅ«nė, Džiuginta² 

[1] Author to whom correspondence should be addressed.;Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, 1870 Frederiksberg C, Denmark

关键词: phage;    fast;    DNA extraction;    spin column;    genome;    sequencing;   

DOI  :  10.3390/mps1030027

学科分类：生物技术

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY