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A rapid competitive method for bacteriophage genomic DNA extraction.

Abbas Soleimani-Delfan¹, Majid Bouzari¹, Ran Wang²

¹Department of Cell and Molecular Biology & Microbiology, Faculty of Biological Science and Technology, University of Isfahan, HezarJereeb Street, 81746-73441, Isfahan, Iran.

Journal of Virological Methods

|April 8, 2021

Summary

A new, rapid, and cost-effective method for bacteriophage (phage) DNA extraction is presented. Acetone precipitation followed by potassium or sodium iodide extraction yields high-quality DNA suitable for genomic analysis in under 10 minutes.

Keywords:

Bacteriophage precipitation DNA extraction Next-generation sequencing Rapid method

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Preparation of Viral DNA from Nucleocapsids

Preparation of Viral DNA from Nucleocapsids

Published on: August 16, 2011

Area of Science:

Microbiology
Molecular Biology
Genomics

Background:

Bacteriophage (phage) DNA extraction is crucial for genomics analysis.
Current methods can be time-consuming and expensive.
A rapid and cost-effective DNA extraction protocol is needed for phage biologists.

Purpose of the Study:

To develop a simple, rapid, and cost-effective method for bacteriophage DNA extraction.
To evaluate different DNA extraction techniques following phage concentration.
To assess the suitability of extracted DNA for downstream genomic applications.

Main Methods:

Phage lysates were concentrated using acetone precipitation.
DNA was extracted using commercial kits, TES lysis buffer, potassium iodide, and sodium iodide.
DNA quality was assessed via agarose gel electrophoresis and UV absorbance (260/280 nm).
Extracted DNA was validated using restriction digestion and next-generation sequencing.

Main Results:

A procedure combining acetone precipitation and potassium or sodium iodide extraction was identified as the most efficient.
This method successfully extracted phage DNA in less than 10 minutes.
The extracted DNA was of high quality, suitable for restriction digestion and next-generation sequencing.

Conclusions:

Acetone precipitation followed by potassium or sodium iodide extraction offers a rapid, cost-effective, and efficient method for bacteriophage DNA extraction.
The protocol yields DNA suitable for various phage genomic analyses, including library preparation and sequencing.
This optimized method benefits phage biologists by significantly reducing processing time and cost.