Molecular Biotechnology

, Volume 15, Issue 2, pp 143–146 | Cite as

Rapid analysis of amplified double-stranded DNA by capillary electrophoresis with laser-induced fluorescence detection

  • Ming-Sun Liu
  • Fu-Tai Albert Chen
Protocol

Abstract

DNA amplification technology has been applied to clinical diagnosis of infectious disease, genetic disorder, and cancer. After in vitro amplification of a particular DNA region, the methods of analysis for these amplified samples play a pivotal role in clinical diagnosis. Conventional gel electrophoresis has been routinely used in the lab for checking DNA. The whole procedure is time consuming and requires more than 1 ng of DNA for detection. To achieve greater performance in DNA diagnosis, we demonstrated capillary electrophoresis with laser induced fluorescence detection for analysis of amplified DNA. The analysis of DNA could be completed within 3 min and the data is directly entered into the computer. Considering the automatic and rapid process, we believe that this method could be routinely utilized for the clinical diagnosis of amplified DNA products.

Index Entries

Capillary electrophoresis double-stranded DNA analysis DNA amplification laser-induced fluorescence detection 

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References

  1. 1.
    Strege, M. and Lagu, A. (1991) Separation of DNA restriction fragments by capillary electrophoresis using coated fused silica capillaries. Anal. Chem. 63, 1233–1236.PubMedCrossRefGoogle Scholar
  2. 2.
    Ulfelder, K. J., Schwartz, H. E., Hall, J. M., and Sunnzeri, F. J. (1992) Restriction fragment length polymorphism analysis of ERBB2 oncogene by capillary electrophoresis. Anal. Biochem. 200, 260–267.PubMedCrossRefGoogle Scholar
  3. 3.
    Schwartz, H. E. and Ulfelder, K. J. (1992) Capillary electrophoresis with laser induced fluorescence detection of PCR fragments thiazole orange. Anal. Chem. 64, 1737–1740.CrossRefGoogle Scholar
  4. 4.
    Rolfs, A., Schuller, L., Finckh, U., Weber-Rolfs, I., eds. (1992) PCR: Clinical Diagnostics and Researeh. Springer-Verlag, Berlin.Google Scholar
  5. 5.
    Liu, M. S., Zang, J., Evangelista, R. A., Rampal, S., and Chen, F. T. A. (1995) Double-stranded DNA analysis by capillary clectrophoresis with laser-induced fluorescence ushig ethidium bromitic as an intercalator. BioTechniques 18, 316–323.PubMedGoogle Scholar
  6. 6.
    Pao, C. C., Kao, S. M., Hor, J. J., and Chang, S. Y. (1993) Lack of mutational alteration in the conserved regions of ZFY and SRY gcne of 46, XY females with gonadal dysgenesis. Hum. Reprod. 8, 224–228.PubMedGoogle Scholar
  7. 7.
    Hixson, J. E. and Vernier, D. T. (1990) Restriction isotyping of human apolipoproteh, E by gene amplification and cleavage with Hhal. J. Lipid. Res. 31, 545–548.PubMedGoogle Scholar

Copyright information

© Humana Press Inc 2000

Authors and Affiliations

  • Ming-Sun Liu
    • 1
  • Fu-Tai Albert Chen
    • 1
  1. 1.Beckman Coulter Inc.Fullerton

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