A robust universal method for extraction of genomic DNA from bacterial species

Sina Atashpaz, Sajjad Khani, Abolfazl Barzegari, Jaleh Barar, Sepideh Zununi Vahed, Reza Azarbaijani, Yadollah Omidi

Research output: Contribution to journalArticlepeer-review

Abstract

The intactness of DNA is the keystone of genome-based clinical investigations, where rapid molecular detection of life-threatening bacteria is largely dependent on the isolation of high-quality DNA. Various protocols have been so far developed for genomic DNA isolation from bacteria, most of which have been claimed to be reproducible with relatively good yields of high-quality DNA. Nonetheless, they are not fully applicable to various types of bacteria, their processing cost is relatively high, and some toxic reagents are used. The routine protocols for DNA extraction appear to be sensitive to species diversity, and may fail to produce high-quality DNA from different species. Such protocols remain time-consuming and tedious, thus to resolve some of these impediments, we report development of a very simple, rapid, and high-throughput protocol for extracting of high-quality DNA from different bacterial species. Based upon our protocol, interfering phenolic compounds were removed from extraction using polyvinylpyrrolidone (PVP) and RNA contamination was precipitated using LiCl. The UV spectrophotometry and gel electrophoresis analysis resulted in high A260/A280 ratio (>1.8) with high intactness of DNA. Subsequent evaluations were performed using some quality-dependent techniques (e.g., RAPD marker and restriction digestions). The isolated DNA from 9 different bacterial species confirmed the accuracy of this protocol which requires no enzymatic processing and accordingly its low-cost making it an appropriate method for large-scale DNA isolation from various bacterial species.

Original languageEnglish
Pages (from-to)538-542
Number of pages5
JournalMicrobiology
Volume79
Issue number4
DOIs
Publication statusPublished - 2010

Keywords

  • bacteria
  • genomic DNA extraction
  • RAPD

ASJC Scopus subject areas

  • Microbiology
  • Applied Microbiology and Biotechnology

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