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

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 extracti...

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Autores principales: Clark, David J, Moore, Catherine M, Flanagan, Marc, Bocxlaer, Katrien Van, Piperaki, Evangelia-Theophano, Yardley, Vanessa, Croft, Simon L, Tyson, John, Whitehouse, Sam P, O’Halloran, Jonathan, Krishna, Sanjeev, Staines, Henry M
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Future Science Ltd 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7115988/
https://www.ncbi.nlm.nih.gov/pubmed/31774301
http://dx.doi.org/10.2144/btn-2019-0086
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author Clark, David J
Moore, Catherine M
Flanagan, Marc
Bocxlaer, Katrien Van
Piperaki, Evangelia-Theophano
Yardley, Vanessa
Croft, Simon L
Tyson, John
Whitehouse, Sam P
O’Halloran, Jonathan
Krishna, Sanjeev
Staines, Henry M
author_facet Clark, David J
Moore, Catherine M
Flanagan, Marc
Bocxlaer, Katrien Van
Piperaki, Evangelia-Theophano
Yardley, Vanessa
Croft, Simon L
Tyson, John
Whitehouse, Sam P
O’Halloran, Jonathan
Krishna, Sanjeev
Staines, Henry M
author_sort Clark, David J
collection PubMed
description 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.
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spelling pubmed-71159882020-08-27 An efficient and novel technology for the extraction of parasite genomic DNA from whole blood or culture Clark, David J Moore, Catherine M Flanagan, Marc Bocxlaer, Katrien Van Piperaki, Evangelia-Theophano Yardley, Vanessa Croft, Simon L Tyson, John Whitehouse, Sam P O’Halloran, Jonathan Krishna, Sanjeev Staines, Henry M Biotechniques Reports 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. Future Science Ltd 2019-11-27 2019-11 /pmc/articles/PMC7115988/ /pubmed/31774301 http://dx.doi.org/10.2144/btn-2019-0086 Text en © 2019 Dr Henry Staines This work is licensed under the Attribution-NonCommercial-NoDerivatives 4.0 Unported License (http://creativecommons.org/licenses/by-nc-nd/4.0/)
spellingShingle Reports
Clark, David J
Moore, Catherine M
Flanagan, Marc
Bocxlaer, Katrien Van
Piperaki, Evangelia-Theophano
Yardley, Vanessa
Croft, Simon L
Tyson, John
Whitehouse, Sam P
O’Halloran, Jonathan
Krishna, Sanjeev
Staines, Henry M
An efficient and novel technology for the extraction of parasite genomic DNA from whole blood or culture
title An efficient and novel technology for the extraction of parasite genomic DNA from whole blood or culture
title_full An efficient and novel technology for the extraction of parasite genomic DNA from whole blood or culture
title_fullStr An efficient and novel technology for the extraction of parasite genomic DNA from whole blood or culture
title_full_unstemmed An efficient and novel technology for the extraction of parasite genomic DNA from whole blood or culture
title_short An efficient and novel technology for the extraction of parasite genomic DNA from whole blood or culture
title_sort efficient and novel technology for the extraction of parasite genomic dna from whole blood or culture
topic Reports
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7115988/
https://www.ncbi.nlm.nih.gov/pubmed/31774301
http://dx.doi.org/10.2144/btn-2019-0086
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