A simple protocol to obtain highly pure Wolbachia endosymbiont DNA for genome sequencing.
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.
School of Biological Sciences, The University of Queensland, Brisbane, Qld 4072, Australia.
Iñaki
I
Iturbe-Ormaetxe
Megan
M
Woolfit
Edwige
E
Rancès
Anne
A
Duplouy
Scott L
SL
O'Neill
D000818:Animals; D009033:Culicidae / Q000382:microbiology; D004269:DNA, Bacterial / Q000302:isolation & purification*; D004175:Diptera / Q000382:microbiology; D016680:Genome, Bacterial; D008967:Molecular Biology / Q000379:methods*; D017422:Sequence Analysis, DNA; D020577:Wolbachia / Q000235:genetics* / Q000302:isolation & purification
D004269:DNA, Bacterial