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A simple protocol to obtain highly pure Wolbachia endosymbiont DNA for genome sequencing
Authors:Iturbe-Ormaetxe Iñaki  Woolfit Megan  Rancès Edwige  Duplouy Anne  O'Neill Scott L
Institution:
  • School of Biological Sciences, The University of Queensland, Brisbane, Qld 4072, Australia
  • 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.
    Keywords:Wolbachia pipientis  Purification  Genomics  Symbiosis
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