Filtration isolation of nucleic acids: A simple and rapid DNA extraction method

Sally M. McFall*, Mário F. Neto, Jennifer L. Reed, Robin L. Wagner

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

FINA, filtration isolation of nucleic acids, is a novel extraction method which utilizes vertical filtration via a separation membrane and absorbent pad to extract cellular DNA from whole blood in less than 2 min. The blood specimen is treated with detergent, mixed briefly and applied by pipet to the separation membrane. The lysate wicks into the blotting pad due to capillary action, capturing the genomic DNA on the surface of the separation membrane. The extracted DNA is retained on the membrane during a simple wash step wherein PCR inhibitors are wicked into the absorbent blotting pad. The membrane containing the entrapped DNA is then added to the PCR reaction without further purification. This simple method does not require laboratory equipment and can be easily implemented with inexpensive laboratory supplies. Here we describe a protocol for highly sensitive detection and quantitation of HIV-1 proviral DNA from 100 μl whole blood as a model for early infant diagnosis of HIV that could readily be adapted to other genetic targets.

Original languageEnglish (US)
Article numbere54289
JournalJournal of Visualized Experiments
Volume2016
Issue number114
DOIs
StatePublished - Aug 6 2016

Keywords

  • DNA extraction
  • Early infant diagnosis
  • HIV
  • Issue 114
  • Molecular biology
  • Molecular diagnostic
  • Paper-based diagnostic
  • Point-of-care
  • Polymerase chain reaction
  • Provirus

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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