Portable lysis apparatus for rapid single-step DNA extraction of Bacillus subtilis

J Appl Microbiol. 2016 Feb;120(2):379-87. doi: 10.1111/jam.13011.

Abstract

Aims: To demonstrate and characterize a portable lysis apparatus for rapid single-step bacterial DNA extraction.

Methods and results: Our portable lysis apparatus employed a novel design consisting of an annular piezo-element with perforated diaphragm. Using Bacillus subtilis as target bacteria, our portable lysis apparatus was able to achieve a normalized percent lysis as high as 66% within 30 s. This is comparable to that by microprobe ultrasonication and almost 7 times higher than that by conventional bead beating. The effect from adding glass beads was predictable. However, the results from the addition of sodium dodecyl sulphate (SDS) were counter-intuitive because a further increase from 0·5 to 1% concentration reduced the lysis performance. The portable lysis apparatus is also at least 1·5-5 times more power efficient than microprobe ultrasonication.

Conclusions: Our portable lysis apparatus is capable of rapidly extracting bacterial DNA and is more power efficient than microprobe ultrasonication. The addition of glass beads or SDS concentration (up to 0·5%) improves its performance.

Significance and impact of the study: The portable lysis apparatus provides a standalone, rapid, low cost and power efficient way of obtaining genomic constituents prior to a variety of bioassays used in the field of environmental, biomedical and other applied microbiology.

Keywords: Bacillus subtilis; bacterial DNA extraction; low cost; portable lysis apparatus; rapid.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Analytic Sample Preparation Methods / instrumentation
  • Analytic Sample Preparation Methods / methods*
  • Bacillus subtilis / chemistry*
  • Bacillus subtilis / genetics
  • Bacillus subtilis / isolation & purification
  • DNA, Bacterial / genetics
  • DNA, Bacterial / isolation & purification*

Substances

  • DNA, Bacterial