Abstract
With the growing applications for the polymerase chain reaction (PCR) in human identity testing, a need exists for more rapid and automated forms of assessing amplification success. Methods that allow the characterization of a sample without consuming much material are helpful, especially in forensic cases. In addition, information regarding the quality and quantity of PCR product can be valuable in some applications. This chapter will focus on quantitating PCR products using capillary electrophoresis (CE). This CE method has proven effective prior to sequence analysis of polymorphisms in the D-loop region of mitochondrial DNA (1).
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Wilson, M. R., Polanskey, D., Butler, J. M., DiZinno, J. A., Replogle, J., Budowle, B. (1995) Extraction, PCR amplification, and sequencing of mitochondrial DNA from human hair shafts. Biotechniques 18, 662–669.
Jenkins, F. J. (1994) Basic methods for the detection of PCR products. PCR Meth. Appl. 4, S77–S82.
Rye, H. S., Dabora, J. M., Quesada, M. A., Mathies, R. A., and Glazer, A. N. (1993) Fluorometric assay using dimeric dyes for double-and single-stranded DNA and RNA with picogram sensitivity. Anal. Biochem. 208, 144–150.
Butler, J. M., McCord, B. R., Jung, J. M., Wilson, M. R., Budowle, B., and Allen, R. O. (1994) Quantitation of polymerase chain reaction products by capillary elec-trophoresis using laser fluorescence. J. Chromatogr. B 658, 271–280.
Schwartz, H. E. and Ulfelder, K. J. (1992) Capillary electrophoresis with laser-induced fluorescence detection of PCR fragments using thiazole orange. Anal. Chem. 64, 1737–1740.
Glazer, A. N. and Rye, H. S. (1992) Stable dye-DNA intercalation complexes as reagents for high-sensitivity fluorescence detection. Nature 359, 859–861.
Lu, W., Han, D. S., Yuan, J., and Andrieu, J. M. (1994) Multi-target PCR analysis by capillary electrophoresis and laser-induced fluorescence. Nature 368, 269–271.
Rossomando, E. F., White, L., and Ulfelder, K. J. (1994) Capillary electrophoresis: separation and quantitation of reverse transcriptase polymerase chain reaction products from polio virus. J. Chromatogr. B 656, 159–168.
Felmlee, T. A., Mitchell, P. S., Ulfelder, K. J., Persing, D. H., and Landers, J. P. (1995) Capillary electrophoresis for the post-amplification detection of a hepatitis C virus-specific DNA product in human serum. J. Cap. Elec. 2(3), 125–130.
Butler, J. M. (1995) Sizing and quantitation of polymerase chain reaction products by capillary electrophoresis for use in DNA typing. Ph. D. dissertation. University of Virginia, Charlottesville.
Hyder, S. M., Hu, C., Needleman, D. S., Sonoda, Y., Wang, X.-Y., and Baker, V. V. (1994) Improved accuracy in direct automated DNA sequencing of small PCR products by optimizing the template concentration. Biotechniques 17, 478–482.
Wilson, M. R., Stoneking, M., Holland, M. M., DiZinno, J. A., and Budowle, B. (1993) Guidelines for the use of mitochondrial DNA sequencing in forensic science. Crime Lab. Dig. 20, 68–77.
Bo>cek, P. and Chrambach, A. (1991) Capillary electrophoresis of DNA in agarose solutions at 40°C. Electrophoresis 12, 1059–1061.
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© 1998 Humana Press Inc., Totowa, NJ
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Butler, J.M. (1998). Rapid Assessment of PCR Product Quality and Quantity by Capillary Electrophoresis. In: Lincoln, P.J., Thomson, J. (eds) Forensic DNA Profiling Protocols. Methods in Molecular Biology, vol 98. Humana Press. https://doi.org/10.1385/0-89603-443-7:39
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DOI: https://doi.org/10.1385/0-89603-443-7:39
Publisher Name: Humana Press
Print ISBN: 978-0-89603-443-3
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