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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...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Portland Press Ltd.
2014
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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. |
format | Online Article Text |
id | pubmed-4206861 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Portland Press Ltd. |
record_format | MEDLINE/PubMed |
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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