4.6 Article

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

Journal

BIOSCIENCE REPORTS
Volume 34, Issue -, Pages 629-634

Publisher

PORTLAND PRESS LTD
DOI: 10.1042/BSR20140105

Keywords

column-based purifications; gDNA extraction; nucleated blood; PCR; salt-precipitation; whole blood

Funding

  1. 2nd Developmental Programme, Joint Council Office, Agency for Science, Technology, and Research, Singapore [1334i0005]
  2. Joint Council Office, Agency for Science, Technology, and Research, Singapore
  3. Quintech Life Sciences Pte Ltd.

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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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