Abstract
The polymerase chain reaction (PCR) is an in vitro method for the amplification of DNA that was introduced in 1985 (1). The principle of the PCR is elegantly simple but the resulting method is extremely powerful. The adoption of the thermostable Taq polymerase in 1988 greatly simplifies the process and enables the automatron of PCR (2). Since then a large number of applications have been developed that are based on the basic PCR theme. The versatility and speed of PCR have revolutionized molecular diagnostics, allowmg the realization of a number of applications that were impossible in the pre-PCR era. This chapter offers an mtroductory guide to the process.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Saiki, R. K., Scharf, S., Faloona, F., Mullis, K. B., Horn, G. T., Erlich, H. A., and Arnheim, N. (1985) Enzymatic amplification of Ī²-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 230, 1350ā1354
Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. J., Higuchi, R., Horn, G. T., Mullis, K. B., and Erlich, H. A. (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239, 487ā491
Yap, E. P. H. and McGee, J. O. (1991) Short PCR product yields improved by lower denaturation temperatures. Nucleic Acids Res 19, 1713.
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, 735lā7367.
Li, H., Gyllensten, U. B., Cui, X., 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, 414ā417.
Hubert, R., MacDonald, M., Gusella, J., and Arnheim, N. (1994) High resolution localization of recombination hot spots using sperm typing. Nut Genet 7, 420ā424.
Handyside, A. H., Lesko, J. G., Tarin, J. J., Winston, R. M., and Hughes, M. R. (1992) Birth of a normal girl after in vitro fertilization and preimplantation diagnostic testing for cystic fibrosis N. Engl. J Med. 327, 905ā909.
Kristjansson, K., Chong, S S., Vandenveyver, I. B., Subramanian, S., Snabes, M. C., and Hughes, M. R. (1994) Preimplantation single cell analyses of dystrophin gene deletions using whole genome amplification. Nat Genet. 6, 19ā23.
Vandenveyver, I. B., Chong, S. S., Cota, J., Bennett, P. R., Fisk, N. M., Handyside, A. H., Cartron, J. P., Le Van Kim, C., Colin, Y., Snabes, M.C., Moise, K. J., and Hughes, M. R. (1995) Single cell analysis of the RhD blood type for use in preimplantation diagnosis in the prevention of severe hemolytic disease of the newborn. Am J Obstet. Gynecol. 172, 533ā540.
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, 1717ā1723.
Birch, D E., Kolmodin, L., Laird, W. J., McKinney, N., Wong, J., and Young, K. K. Y. (1996) Simplified Hot Start PCR. Nature 381, 445,446.
Eckert, K. A. and Kunkel, T. A. DNA polymerase fidelity and the polymerase chain reaction. (1991) PCR Methods Appl. 1, 17ā24.
Cariello, N. F., Swenberg, J. A., and Skopek, T R. (1991) Fidelity of Thermococcus litorahs DNA polymerase (Vent) in PCR determined by denaturing gradient gel-electrophoresis. Nucleic Acids Res. 19, 4193ā4198.
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, 6230ā6234.
Sutcharitchan, P., Saiki, R, Huisman, T., Kutlar, A., Mckie, V., and Erlich, H (1995) Reverse dot-blot detection of the African-American beta-thalassemia mutations. Blood 86, 1580ā1585
Rady, M., Dalcamo, E., Seia, M., Iapichino, L., Ferari, M., Russo, S., Romeo, G., and Maggie, A (1995) Simultaneous detection of 14 Italian cystic-fibrosis mutations in 7 exons by reverse dot-blot analysis. Mol Cell. Probes 9, 357ā360.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
Ā© 1998 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Dennis Lo, Y.M. (1998). Introduction to the Polymerase Chain Reaction. In: Lo, Y.M.D. (eds) Clinical Applications of PCR. Methods in Molecular Medicineā¢, vol 16. Humana Press. https://doi.org/10.1385/0-89603-499-2:3
Download citation
DOI: https://doi.org/10.1385/0-89603-499-2:3
Publisher Name: Humana Press
Print ISBN: 978-0-89603-499-0
Online ISBN: 978-1-59259-600-3
eBook Packages: Springer Protocols