Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2018 Jul 27;1(3):27.
doi: 10.3390/mps1030027.

A Rapid Bacteriophage DNA Extraction Method

Džiuginta Jakočiūnė  1 , Arshnee Moodley  2
Affiliations

A Rapid Bacteriophage DNA Extraction Method

Džiuginta Jakočiūnė et al. Methods Protoc. .

Abstract

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.

Keywords: DNA extraction; fast; genome; phage; sequencing; spin column.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Five µL of extracted DNA was loaded on 1% agarose gel. L—GeneRuler 1 kb DNA Ladder (250–10,000 bp, Thermo Scientific, Roskilde, Denmark); S13–S16 phage DNA.
See this image and copyright information in PMC

References

    1. Sulakvelidze A. Safety by nature: Potential bacteriophage applications. Microbe Mag. 2011;6:122–126. doi: 10.1128/microbe.6.122.1. - DOI
    1. Vandenheuvel D., Lavigne R., Ussow H.B. Bacteriophage therapy: Advances in formulation strategies and human clinical trials. Annu. Rev. Virol. 2015;2:599–618. doi: 10.1146/annurev-virology-100114-054915. - DOI - PubMed
    1. Pickard D.J.J. Preparation of Bacteriophage Lysates and Pure DNA. In: Clokie M.R.J., Kropinski A.M., editors. Bacteriophages. Volume 2. Humana Press; New York, NY, USA: 2009. pp. 3–10. Methods in Molecular Biology. - PubMed
    1. Sambrook J., Russell D.W. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press; Cold Spring Harbor, NY, USA: 2001.
    1. Kropinski A.M., Mazzocco A., Waddell T.E., Lingohr E., Johnson R.P. Enumeration of Bacteriophages by Double Agar Overlay Plaque Assay. In: Clokie M.R.J., Kropinski A.M., editors. Bacteriophages. Volume 1. Humana Press; New York, NY, USA: 2009. pp. 69–76. - PubMed

LinkOut - more resources