Elsevier

Analytical Biochemistry

Volume 283, Issue 2, 1 August 2000, Pages 175-191
Analytical Biochemistry

Regular Article
Evaluation of Silica Resins for Direct and Efficient Extraction of DNA from Complex Biological Matrices in a Miniaturized Format

https://doi.org/10.1006/abio.2000.4577Get rights and content

Abstract

For DNA purification to be functionally integrated into the microchip for high-throughput DNA analysis, a miniaturized purification process must be developed that can be easily adapted to the microchip format. In this study, we evaluate the effectiveness of a variety of silica resins for miniaturized DNA purification and gauge the potential usefulness for on-chip solid-phase extraction. A micro-solid-phase extraction (μSPE) device containing only nanograms of silica resin is shown to be effective for the adsorption and desorption of DNA in the picogram–nanogram mass range. Fluorescence spectroscopy as well as capillary electrophoresis with laser-induced fluorescence detection is employed for the analysis of DNA recovered from solid-phase resins, while the polymerase chain reaction (PCR) is used to evaluate the amplifiable nature of the eluted DNA. We demonstrate that DNA can be directly recovered from white blood cells with an efficiency of roughly 70%, while greater than 80% of the protein is removed with a 500-nl bed volume μSPE process that takes less than 10 min. With a capacity in the range of 10–30 ng/mg of silica resin, we show that the DNA extracted from white blood cells, cultured cancer cells, and even whole blood on the low microliter scale is suitable for direct PCR amplification. The miniaturized format as well as rapid time frame for DNA extraction is compatible with the fast electrophoresis on microfabricated chips.

References (55)

  • H. Tian et al.

    Genomics

    (2000)
  • J. Cheng et al.

    Anal. Biochem.

    (1998)
  • P. Wilding et al.

    Anal. Biochem.

    (1998)
  • M.A. Marko et al.

    Anal. Biochem.

    (1982)
  • E. Matitashvili et al.

    Anal. Biochem.

    (1997)
  • K. Wang et al.

    Anal. Biochem.

    (1995)
  • M.A. Strausbauch et al.

    J. Chromatogr.

    (1995)
  • M.E. Roche et al.

    Anal. Biochem.

    (1998)
  • V.L. Singer et al.

    Anal. Biochem.

    (1997)
  • K.A. Melzak et al.

    J. Colloid Interface Sci.

    (1996)
  • K. Unger et al.

    J. Chromatogr.

    (1976)
  • I. Kheterpal et al.

    Anal. Chem.

    (1999)
  • Q. Gao et al.

    Electrophoresis

    (1999)
  • Y. Wang et al.

    Anal. Chem.

    (1995)
  • Y. Wang et al.

    Electrophoresis

    (1996)
  • W.T. Hofgärtner et al.

    Clin. Chem.

    (1999)
  • A.T. Woolley et al.

    Anal. Chem.

    (1997)
  • S. Liu et al.

    Anal. Chem.

    (1999)
  • Y. Shi et al.

    Anal. Chem.

    (1999)
  • P.C. Simpson et al.

    Proc. Natl. Acad. Sci. USA

    (1998)
  • N.J. Munro et al.

    Clin. Chem.

    (1999)
  • Tian, H, Brody, L. C, Munro, N. J, and, Landers, J. P. 2000, Rapid detection of deletion, insertion and substitution...
  • R. Elles

    Molecular Diagnosis of Genetic Disease

    (1996)
  • R.P. Oda et al.

    Anal. Chem.

    (1998)
  • Hühmer, A. F. R, and, Landers, J. P. 2000,...
  • A.T. Woolley et al.

    Anal. Chem.

    (1996)
  • L.C. Waters et al.

    Anal. Chem.

    (1998)
  • Cited by (241)

    • Materials and methods for microfabrication of microfluidic devices

      2021, Microfluidic Devices for Biomedical Applications
    • Protein-based bionanocomposites

      2020, Bionanocomposites: Green Synthesis and Applications
    View all citing articles on Scopus
    1

    Current address: Texas Instruments, Inc., Analytical Sensor Group, 13536 N. Central Expressway, MS945, Dallas, TX 75243. E-mail: [email protected].

    2

    To whom correspondence should be addressed at Department of Chemistry, McCormick Road, University of Virginia, Charlottesville, VA 22901. Fax: 412-243-8852. E-mail: [email protected].

    View full text