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Introduction to the Polymerase Chain Reaction

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Book cover Clinical Applications of PCR

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 336))

Abstract

The polymerase chain reaction (PCR) is an in vitro method for the amplification of DNA. Since the introduction of the PCR in 1985, it has become an indispensable technique for many applications in scientific research and clinical and forensic investigations. In this chapter, the principle and setup of PCR, as well as the methods for analyzing PCR products, will be discussed.

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References

  1. Saiki, R. K., Scharf, S., Faloona, F., et al. (1985) Enzymatic amplification of betaglobin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 230,1350–1354.

    Article  PubMed  CAS  Google Scholar 

  2. Saiki, R. K., Gelfand, D. H., Stoffel, S., et al. (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase.Science 239,487–491.

    Article  PubMed  CAS  Google Scholar 

  3. Nagai, H., Murakami, Y., Morita, Y., Yokoyama, K., and Tamiya, E. (2001) Development of a microchamber array for picoliter PCR.Anal. Chem. 73, 1043–1047.

    Article  PubMed  CAS  Google Scholar 

  4. Puppe, W., Weigl, J. A., Aron, G., et al. (2004) Evaluation of a multiplex reverse ranscriptase PCR ELISA for the detection of nine respiratory tract pathogens. J. Clin. Virol. 30, 165–174.

    Article  PubMed  CAS  Google Scholar 

  5. Tournier, I., Paillerets, B. B., Sobol H., et al. (2004) Significant contribution of germline BRCA2 rearrangements in male breast cancer families. Cancer Res. 64, 8143–8147.

    Article  PubMed  CAS  Google Scholar 

  6. Yap, E. P. and McGee, J. O. (1991) Short PCR product yields improved by lower denaturation temperatures. Nucleic Acids Res. 19, 1713.

    Article  PubMed  CAS  Google Scholar 

  7. Higuchi, R., Krummel, B., and Saiki, R. K. (1988) A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions. Nucleic Acids Res. 16, 7351–7367.

    Article  PubMed  CAS  Google Scholar 

  8. Li, H. H., Gyllensten U. B., Cui X. F., Saiki R. K., Erlich H. A., and Arnheim N. (1988) Amplification and analysis of DNA sequences in single human sperm and diploid cells. Nature 335, 414–417.

    Article  PubMed  CAS  Google Scholar 

  9. Jeffreys, A. J., Kauppi, L., and Neumann, R. (2001) Intensely punctate meiotic recombination in the class II region of the major histocompatibility complex. Nat. Genet. 29, 217–222.

    Article  PubMed  CAS  Google Scholar 

  10. Moutou, C., Gardes, N., and Viville, S. (2004) New tools for preimplantation genetic diagnosis of Huntington’s disease and their clinical applications. Eur. J. Hum. Genet. 12, 1007–1014.

    Article  PubMed  CAS  Google Scholar 

  11. Chou, Q., Russell, M., Birch, D. E., Raymond, J., and Bloch, W. (1992) Prevention of pre-PCR mis-priming and primer dimerization improves low-copy-number amplifications. Nucleic Acids Res. 20, 1717–1723.

    Article  PubMed  CAS  Google Scholar 

  12. Birch, D. E. (1996) Simplified hot start PCR. Nature 381, 445–446.

    Article  PubMed  CAS  Google Scholar 

  13. Eckert, K. A. and Kunkel, T. A. (1991) DNA polymerase fidelity and the polymerase chain reaction. PCR Methods Appl. 1, 17–24.

    PubMed  CAS  Google Scholar 

  14. Cariello, N. F., Swenberg, J. A., and Skopek, T. R. (1991) Fidelity of Thermococcus litoralis DNA polymerase (Vent) in PCR determined by denaturing gradient gel electrophoresis. Nucleic Acids Res. 19, 4193–4198.

    Article  PubMed  CAS  Google Scholar 

  15. Krishnan, M., Ugaz, V. M., and Burns, M. A. (2002) PCR in a Rayleigh-Benard convection cell. Science 298, 793.

    Article  PubMed  Google Scholar 

  16. Wheeler, E. K., Benett, W., Stratton, P., et al. (2004) Convectively driven polymerase chain reaction thermal cycler. Anal. Chem. 76, 4011–4016.

    Article  PubMed  CAS  Google Scholar 

  17. Braun, D., Goddard, N. L., and Libchaber, A. (2003) Exponential DNA replication by laminar convection. Phys. Rev. Lett. 91, 158103.

    Article  PubMed  Google Scholar 

  18. Saiki, R. K., Walsh, P. S., Levenson, C. H., and Erlich, H. A. (1989) Genetic analysis of amplified DNA with immobilized sequence-specific oligonucleotide probes. Proc. Natl. Acad. Sci. USA 86, 6230–6234.

    Article  PubMed  CAS  Google Scholar 

  19. Dhami, P., Coffey, A. J., Abbs, S., et al. (2005) Exon array CGH: detection of copy-number changes at the resolution of individual exons in the human genome. Am. J. Hum. Genet. 76, 750–762.

    Article  PubMed  CAS  Google Scholar 

  20. Devries, S., Nyante, S., Korkola, J., et al. (2005) Array-based comparative genomic hybridization from formalin-fixed, paraffin-embedded breast tumors. J. Mol. Diagn. 7, 65–71.

    Article  PubMed  CAS  Google Scholar 

  21. Hu, D. G., Webb, G., and Hussey, N. (2004) Aneuploidy detection in single cells using DNA array-based comparative genomic hybridization. Mol. Hum. Reprod. 10, 283–289.

    Article  PubMed  CAS  Google Scholar 

  22. Heid, C. A., Stevens, J., Livak, K. J., and Williams, P. M. (1996) Real time quantitative PCR. Genome Res. 6, 986–994.

    Article  PubMed  CAS  Google Scholar 

  23. Nelson, M. R., Marnellos, G., Kammerer, S., et al. (2004) Large-scale validation of single nucleotide polymorphisms in gene regions. Genome Res. 14, 1664–1668.

    Article  PubMed  CAS  Google Scholar 

  24. Buetow, K. H., Edmonson, M., MacDonald, R., et al. (2001) High-throughput development and characterization of a genomewide collection of gene-based single nucleotide polymorphism markers by chip-based matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry. Proc. Natl. Acad. Sci. USA 98, 581–584.

    Article  PubMed  CAS  Google Scholar 

  25. Mohlke, K. L., Erdos, M. R., Scott, L. J., et al. (2002) High-throughput screening for evidence of association by using mass spectrometry genotyping on DNA pools. Proc. Natl. Acad. Sci. USA 99, 16,928–16,933.

    Article  PubMed  CAS  Google Scholar 

  26. Lleonart, M. E., Ramon y Cajal, S., Groopman, J. D., and Friesen, M. D. (2004) Sensitive and specific detection of K-ras mutations in colon tumors by short oligonucleotide mass analysis. Nucleic Acids Res. 32, e53.

    Article  PubMed  Google Scholar 

  27. Ding, C., Chiu, R. W. K., Lau, T. K., et al. (2004) MS analysis of single-nucleotide differences in circulating nucleic acids: Application to noninvasive prenatal diagnosis. Proc. Natl. Acad. Sci. USA 101, 10,762–10,767.

    Article  PubMed  CAS  Google Scholar 

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Lo, Y.M.D., Chan, K.C.A. (2006). Introduction to the Polymerase Chain Reaction. In: Lo, Y.M.D., Chiu, R.W.K., Chan, K.C.A. (eds) Clinical Applications of PCR. Methods in Molecular Biology™, vol 336. Humana Press. https://doi.org/10.1385/1-59745-074-X:1

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  • DOI: https://doi.org/10.1385/1-59745-074-X:1

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-348-0

  • Online ISBN: 978-1-59745-074-4

  • eBook Packages: Springer Protocols

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