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Comparison of customized spin-column and salt-precipitation finger-prick blood DNA extraction

gDNA (genomic DNA extraction from blood is a fundamental process in many diagnostic, identification and research applications. Numerous extraction methods have been reported and are available commercially. However, there is insufficient understanding of the impact of chemical buffers on DNA yield fr...

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Detalles Bibliográficos
Autores principales: Poh, Jun-Jie, Gan, Samuel Ken-En
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Portland Press Ltd. 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4206861/
https://www.ncbi.nlm.nih.gov/pubmed/25222694
http://dx.doi.org/10.1042/BSR20140105
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author Poh, Jun-Jie
Gan, Samuel Ken-En
author_facet Poh, Jun-Jie
Gan, Samuel Ken-En
author_sort Poh, Jun-Jie
collection PubMed
description gDNA (genomic DNA extraction from blood is a fundamental process in many diagnostic, identification and research applications. Numerous extraction methods have been reported and are available commercially. However, there is insufficient understanding of the impact of chemical buffers on DNA yield from either whole or nucleated blood. Moreover, these commercial kits are often costly, constraining less well-funded laboratories to traditional and more cost-effective salt-precipitation methods. Towards this, we compared a salt-precipitation and a customized cost-effective spin-column-based method, studying the impact of different chemical constituents on the yields. This customized method resulted in a shortening of the extraction process, higher gDNA yields, and more successful PCR amplification of gDNA genes compared with the salt-precipitation method. Optimizing different chemical buffers on whole- and nucleated blood materials further revealed that certain chemicals boosted extractions from whole- but not nucleated blood. These findings may be useful to laboratories that do not have ready access to commercial kits, and improve their nucleic acid extractions from blood economically.
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spelling pubmed-42068612014-10-27 Comparison of customized spin-column and salt-precipitation finger-prick blood DNA extraction Poh, Jun-Jie Gan, Samuel Ken-En Biosci Rep Original Paper gDNA (genomic DNA extraction from blood is a fundamental process in many diagnostic, identification and research applications. Numerous extraction methods have been reported and are available commercially. However, there is insufficient understanding of the impact of chemical buffers on DNA yield from either whole or nucleated blood. Moreover, these commercial kits are often costly, constraining less well-funded laboratories to traditional and more cost-effective salt-precipitation methods. Towards this, we compared a salt-precipitation and a customized cost-effective spin-column-based method, studying the impact of different chemical constituents on the yields. This customized method resulted in a shortening of the extraction process, higher gDNA yields, and more successful PCR amplification of gDNA genes compared with the salt-precipitation method. Optimizing different chemical buffers on whole- and nucleated blood materials further revealed that certain chemicals boosted extractions from whole- but not nucleated blood. These findings may be useful to laboratories that do not have ready access to commercial kits, and improve their nucleic acid extractions from blood economically. Portland Press Ltd. 2014-10-22 /pmc/articles/PMC4206861/ /pubmed/25222694 http://dx.doi.org/10.1042/BSR20140105 Text en © 2014 The Author(s) This is an Open Access article distributed under the terms of the Creative Commons Attribution Licence (CC-BY) (http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited. http://creativecommons.org/licenses/by/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Licence (CC-BY) (http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Paper
Poh, Jun-Jie
Gan, Samuel Ken-En
Comparison of customized spin-column and salt-precipitation finger-prick blood DNA extraction
title Comparison of customized spin-column and salt-precipitation finger-prick blood DNA extraction
title_full Comparison of customized spin-column and salt-precipitation finger-prick blood DNA extraction
title_fullStr Comparison of customized spin-column and salt-precipitation finger-prick blood DNA extraction
title_full_unstemmed Comparison of customized spin-column and salt-precipitation finger-prick blood DNA extraction
title_short Comparison of customized spin-column and salt-precipitation finger-prick blood DNA extraction
title_sort comparison of customized spin-column and salt-precipitation finger-prick blood dna extraction
topic Original Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4206861/
https://www.ncbi.nlm.nih.gov/pubmed/25222694
http://dx.doi.org/10.1042/BSR20140105
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