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An efficient and novel technology for the extraction of parasite genomic DNA from whole blood or culture.

David J ClarkCatherine M MooreMarc FlanaganKatrien Van BocxlaerEvangelia-Theophano PiperakiVanessa YardleySimon L CroftJohn TysonSam P WhitehouseJonathan O'HalloranSanjeev KrishnaHenry M Staines
Published in: BioTechniques (2019)
The aim of this study was to assess pathogen DNA extraction with a new spin column-based method (DNA-XT). DNA from either whole-blood samples spiked with Plasmodium falciparum or Leishmania donovani amastigote culture was extracted with DNA-XT and compared with that produced by a commercial extraction kit (DNeasy®). Eluates from large and small sample volumes were assessed by PCR and spectroscopy. Using a small volume (5 μl) of blood, the DNA-XT and DNeasy methods produced eluates with similar DNA concentrations (0.63 vs 1.06 ng/μl, respectively). The DNA-XT method produced DNA with lower PCR inhibition than DNeasy. The new technique was also twice as fast and required fewer plastics and manipulations but had reduced total recovered DNA compared with DNeasy.
Keyphrases
  • circulating tumor
  • single molecule
  • cell free
  • nucleic acid
  • plasmodium falciparum
  • high resolution
  • density functional theory