Jakočiūnė Džiuginta, Moodley Arshnee
Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, 1870 Frederiksberg C, Denmark.
Methods Protoc. 2018 Jul 27;1(3):27. doi: 10.3390/mps1030027.
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 × 10 plaque-forming units (PFU)/mL. This DNA was suitable for library preparation using the Nextera XT kit and sequencing on the Illumina MiSeq platform.
噬菌体作为非抗生素替代品，被广泛研究以规避抗生素耐药性的产生，同时也是对抗耐药菌的最后手段治疗选择。为了更好地了解噬菌体并确定其是否携带潜在毒力因子，人们采用了全基因组测序技术，而这需要高质量的噬菌体DNA。传统的噬菌体DNA提取方法繁琐且耗时，需要特殊设备，如超速离心机。在此，我们描述了一种快速简便的方法（四小时内），用于从滴度高于1.0×10噬菌斑形成单位（PFU）/mL的双链DNA（dsDNA）噬菌体中提取DNA。该DNA适用于使用Nextera XT试剂盒制备文库，并在Illumina MiSeq平台上进行测序。
