A simple protocol to obtain highly pure Wolbachia endosymbiont DNA for genome sequencing

Iñaki Iturbe-Ormaetxe; Megan Woolfit; Edwige Rancès; Anne Duplouy; Scott L O'Neill

doi:10.1016/j.mimet.2010.10.019

A simple protocol to obtain highly pure Wolbachia endosymbiont DNA for genome sequencing

J Microbiol Methods. 2011 Jan;84(1):134-6. doi: 10.1016/j.mimet.2010.10.019. Epub 2010 Oct 31.

Authors

Iñaki Iturbe-Ormaetxe¹, Megan Woolfit, Edwige Rancès, Anne Duplouy, Scott L O'Neill

Affiliation

¹ School of Biological Sciences, The University of Queensland, Brisbane, Qld 4072, Australia.

PMID: 21047535
DOI: 10.1016/j.mimet.2010.10.019

Abstract

Most genome sequencing projects using intracellular bacteria face difficulties in obtaining sufficient bacterial DNA free of host contamination. We have developed a simple and rapid protocol to isolate endosymbiont DNA virtually free from fly and mosquito host DNA. We purified DNA from six Wolbachia strains in preparation for genome sequencing using this method, and achieved up to 97% pure Wolbachia sequence, even after using frozen insects. This is a significant improvement for future Wolbachia and other endosymbiont genome projects.

MeSH terms

Animals
Culicidae / microbiology
DNA, Bacterial / isolation & purification*
Diptera / microbiology
Genome, Bacterial*
Molecular Biology / methods*
Sequence Analysis, DNA
Wolbachia / genetics*
Wolbachia / isolation & purification

Substances

DNA, Bacterial