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Purification of DNA/RNA in a Microfluidic Device

  • Andy Fan
  • Samantha Byrnes
  • Catherine KlapperichEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 949)

Abstract

Often, modern diagnostic techniques require the isolation and purification of nucleic acids directly from patient samples such as blood or stool. Many diagnostic tests are being miniaturized onto micro-sized platforms and integrated into microfluidic devices due to the economies resulting from smaller sample and reagent volumes. Often, these devices perform sample preparation in series with the diagnostic tests. The sample preparation steps are vital in order to purify the desired genetic material from potential inhibitors that can interfere with the outcome of the test. There are various techniques used to selectively capture the nucleic acids while washing away potential contamination (proteins, enzymes, lipids, etc.). Two of the most common forms of selective capture are based on nucleic acid binding to silica surface or on the precipitation of nucleic acids with or without the presence of a carrier species. Each of these methods can be performed in liquid phase or in a solid support such as an extraction column. Here we discuss both methods and address microfluidic applications.

Key words

Sample preparation Microfluidic device Diagnostic tests Micro-solid phase extraction Purification of DNA/RNA Blood lysis 

References

  1. 1.
    Boom R, Sol CJA, Salimans MMM, Jansen CL et al (1990) Rapid and simple method for purification of nucleic acids. J Clin Microbiol 28:495–503Google Scholar
  2. 2.
    Milton ID, Carter MJ (1993) An Inexpensive and simple method for DNA purifications on silica particles. Nucleic Acids Res 21(4):1044CrossRefGoogle Scholar
  3. 3.
    Zhao L, Fu ZF (2009) Isolation of viral RNA from cultures, chapter 3. In: Liu D (ed) Handbook of nucleic acid purification. CRC, Boca Raton, FL, pp 41–60Google Scholar
  4. 4.
    Bhattacharyya A, Klapperich CM (2006) Thermoplastic microfluidic device for on-chip purification of nucleic acids of disposable diagnostics. Anal Chem 78:788–792CrossRefGoogle Scholar
  5. 5.
    Wolfe KA, Breadmore MC, Ferrance JP et al (2002) Toward a microchip-based solid-phase extraction method for isolation of nucleic acids. Electrophoresis 23:727–733CrossRefGoogle Scholar
  6. 6.
    Tan A, Benetton S, Henion JD (2003) Chip-based solid-phase extraction pretreatment for direct electrospray mass spectrometry analysis using an array of monolithic columns in a polymeric substrate. Anal Chem 75:5504–5511CrossRefGoogle Scholar
  7. 7.
    Han S-I, Han K-H, Frazier AB, Ferrance JP, Landers JP (2009) An automated micro-solid phase extraction device involving integrated high-pressure microvalves for genetic sample preparation. Biomed Microdevices 11:935–942CrossRefGoogle Scholar
  8. 8.
    Sambrook J, Russell DW (2001) Extraction, purification, and analysis of mRNA from eukaryotic cells, molecular cloning: a laboratory manual, chapter 7, 3rd edn. Cold Springs Harbor Laboratory Press, Cold Springs Harbor, NYGoogle Scholar
  9. 9.
    Hanson LA (2009) Isolation of viral DNA from cultures, chapter 2. In: Liu D (ed) Handbook of nucleic acid purification. Boca Raton, FL, CRC, pp 23–39Google Scholar
  10. 10.
    Breadmore MC, Wolfe KA, Arcibal IG et al (2003) Microchip-based purification of DNA from biological samples. Anal Chem 75:1800–1886CrossRefGoogle Scholar
  11. 11.
    Chatterjee A, Mirer PL, Santamaria EZ, Klapperich C, Sharon A, Sauer-Budge AF (2010) RNA isolation from mammalian cells using porous polymer monoliths: an approach for high-throughput automation. Anal Chem 82:4344–4356CrossRefGoogle Scholar
  12. 12.
    Alexandrov A, Dutta K, Pascal SM (2001) Sensitive ribonuclease protection assay employing glycogen as a carrier and single inactivation/precipitation step. Biotechniques 30:1198–1204Google Scholar
  13. 13.
    Chomczynski P, Sacchi N (2006) The single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction: twenty-something years on. Nat Protoc 1:581–585CrossRefGoogle Scholar
  14. 14.
    Giordano BC, Ferrance J, Swedberg S et al (2001) Polymerase chain reaction in polymeric microchips: DNA amplification in less than 240 seconds. Anal Biochem 291:124–132CrossRefGoogle Scholar
  15. 15.
    Rohr T, Olgetree DF, Svec F, Frechet JM (2003) Surface functionalization of thermoplastic polymers for the fabrication of microfluidic devices by photoinitiated grafting. Adv Funct Mater 13:264–270CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media,LLC 2013

Authors and Affiliations

  • Andy Fan
    • 1
  • Samantha Byrnes
    • 1
  • Catherine Klapperich
    • 1
    Email author
  1. 1.Department of Biomedical EngineeringBoston UniversityBostonUSA

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