Literature DB >> 27583575

Filtration Isolation of Nucleic Acids: A Simple and Rapid DNA Extraction Method.

Sally M McFall1, Mário F Neto2, Jennifer L Reed2, Robin L Wagner3.   

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.

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Year:  2016        PMID: 27583575      PMCID: PMC5091753          DOI: 10.3791/54289

Source DB:  PubMed          Journal:  J Vis Exp        ISSN: 1940-087X            Impact factor:   1.355


  16 in total

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Journal:  Mol Biotechnol       Date:  2004-02       Impact factor: 2.695

Review 2.  Diagnostics for the developing world.

Authors:  David Mabey; Rosanna W Peeling; Andrew Ustianowski; Mark D Perkins
Journal:  Nat Rev Microbiol       Date:  2004-03       Impact factor: 60.633

3.  Rapid and simple method for purification of nucleic acids.

Authors:  R Boom; C J Sol; M M Salimans; C L Jansen; P M Wertheim-van Dillen; J van der Noordaa
Journal:  J Clin Microbiol       Date:  1990-03       Impact factor: 5.948

Review 4.  Rapid tests for sexually transmitted infections (STIs): the way forward.

Authors:  R W Peeling; K K Holmes; D Mabey; A Ronald
Journal:  Sex Transm Infect       Date:  2006-12-06       Impact factor: 3.519

Review 5.  PCR inhibitors - occurrence, properties and removal.

Authors:  C Schrader; A Schielke; L Ellerbroek; R Johne
Journal:  J Appl Microbiol       Date:  2012-07-24       Impact factor: 3.772

6.  One-step purification and concentration of DNA in porous membranes for point-of-care applications.

Authors:  S A Byrnes; J D Bishop; L Lafleur; J R Buser; B Lutz; P Yager
Journal:  Lab Chip       Date:  2015-06-21       Impact factor: 6.799

7.  A simple and rapid DNA extraction method from whole blood for highly sensitive detection and quantitation of HIV-1 proviral DNA by real-time PCR.

Authors:  Sally M McFall; Robin L Wagner; Sujit R Jangam; Douglas H Yamada; Diana Hardie; David M Kelso
Journal:  J Virol Methods       Date:  2015-02-12       Impact factor: 2.014

8.  "Paper Machine" for Molecular Diagnostics.

Authors:  John T Connelly; Jason P Rolland; George M Whitesides
Journal:  Anal Chem       Date:  2015-07-13       Impact factor: 6.986

9.  Early antiretroviral therapy and mortality among HIV-infected infants.

Authors:  Avy Violari; Mark F Cotton; Diana M Gibb; Abdel G Babiker; Jan Steyn; Shabir A Madhi; Patrick Jean-Philippe; James A McIntyre
Journal:  N Engl J Med       Date:  2008-11-20       Impact factor: 91.245

10.  Dried blood spots--preparing and processing for use in immunoassays and in molecular techniques.

Authors:  Nico Grüner; Oumaima Stambouli; R Stefan Ross
Journal:  J Vis Exp       Date:  2015-03-13       Impact factor: 1.355

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  1 in total

1.  A simple and rapid DNA extraction method for Chlamydia trachomatis detection from urogenital swabs.

Authors:  Matthew A Butzler; Jennifer L Reed; Sally M McFall
Journal:  Diagn Microbiol Infect Dis       Date:  2017-08-17       Impact factor: 2.803

  1 in total

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