Polymerase Chain Reaction

  • Beverly C. Delidow
  • John P. Lynch
  • John J. Peluso
  • Bruce A. White
Part of the Methods in Molecular Biology™ book series (MIMB, volume 58)


The melding of a technique for repeated rounds of DNA synthesis with the discovery of a thermostable DNA polymerase has given scientists the very powerful technique known as polymerase chain reaction (PCR). PCR is based on three simple steps required for any DNA synthesis reaction: (1) denaturation of the template into single strands; (2) annealing of primers to each original strand for new strand synthesis; and (3) extension of the new DNA strands from the primers. These reactions may be carried out with any DNA polymerase and result in the synthesis of defined portions of the original DNA sequence. However, in order to achieve more than one round of synthesis, the templates must again be denatured, which requires temperatures well above those that inactivate most enzymes. Therefore, initial attempts at cyclic DNA synthesis were carried out by adding fresh polymerase after each denaturation step (1,2). The cost of such a protocol becomes rapidly prohibitive.


Polymerase Chain Reaction Polymerase Chain Reaction Product Microfuge Tube DEPC Water Molecular Biology Grade 
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  1. 1.
    Saiki, R., Scharf, S., Faloona, F., Mullis, K. B., Horn, G. T., Erlich, H. A., and Arnheim, N. (1985) Enzymatic amplification of β-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 230, 1350–1354.PubMedCrossRefGoogle Scholar
  2. 2.
    Mullis, K. B. and Faloona, F. A. (1987) Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods Enzymol. 155, 335–350.PubMedCrossRefGoogle Scholar
  3. 3.
    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, 487–491.PubMedCrossRefGoogle Scholar
  4. 4.
    Erlich, H. A., Gelfand, D., and Sninsky, J. J. (1991) Recent advances in the polymerase chain reaction. Science 252, 1643–1651.PubMedCrossRefGoogle Scholar
  5. 5.
    Robert, S. S. (1991) Amplification of nucleic acid sequences: the choices multiply. J. NIH Res. 3(2), 81–94.Google Scholar
  6. 6.
    Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.Google Scholar
  7. 7.
    Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Smith, J. A., Seidman, J. G., and Struhl, K. (eds.) (1987) Current Protocols in Molecular Biology. Wiley Interscience, New York.Google Scholar
  8. 8.
    Dycaico, M. and Mather, S. (1991) Reduce PCR false positives using the Stratalinker UV crosslinker. Stratagene Strategies 4(3), 39, 40.Google Scholar
  9. 9.
    Sarkar, G. and Sommer, S. S. (1990) Shedding light on PCR contamination. Nature 343, 27.PubMedCrossRefGoogle Scholar
  10. 10.
    Zintz, C. B. and Beebe, D. C. (1991) Rapid re-amplification of PCR products purified from low melting point agarose gels. Biotechniques 11, 158–162.PubMedGoogle Scholar
  11. 11.
    Delidow, B. C., Peluso, J. J. and White, B. A. (1989) Quantitative measurement of mRNAs by polymerase chain reaction. Gene Anal. Tech. 6, 120–124.PubMedCrossRefGoogle Scholar
  12. 12.
    Chomczynski, P. and Sacchi, N. (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem. 162, 156–159.PubMedCrossRefGoogle Scholar
  13. 13.
    Chomczynski, P. (1991) Isolation of RNA by the RNAzol method. TEL-Test, Inc., Bull. 2, 1–5.Google Scholar
  14. 14.
    Myers, T. W. and Gelfand, D. H. (1991) Reverse transcription and DNA amplification by a Thermus thermophilus DNA polymerase. Biochemistry 30, 7661–7666.PubMedCrossRefGoogle Scholar
  15. 15.
    Zimmer, A. and Gruss, P. (1991) Use of polymerase chain reaction (PCR) to detect homologous recombination in transfected cell lines, in Methods in Molecular Biology, vol 7: Gene Transfer and Expression Protocols (Murray, E. J., ed.) Humana, Clifton, NJ, pp. 411–418.CrossRefGoogle Scholar
  16. 16.
    Ohara, O., Dorit, R. L., and Gilbert, W. (1989) One-sided polymerase chain reaction: the amplification of cDNA. Proc. Natl. Acad. Sci. USA 86, 5673–5677.PubMedCrossRefGoogle Scholar
  17. 17.
    Belyavsky, A. (1989) Polymerase chant reaction in the presence of NuSieve™ GTG agarose. FMC Resolutions 5, 1,2.Google Scholar
  18. 18.
    Belyavsky, A., Vinogradova, T., and Rajewsky, K. (1989) PCR-based cDNA library construction: general cDNA libraries at the level of a few cells. Nucleic Acids Res. 17, 2919–2932.PubMedCrossRefGoogle Scholar
  19. 19.
    Delidow, B. C., White, B. A., and Peluso, J. J. (1990) Gonadotropin induction of c-fos and c-myc expression and deoxyribonucleic acid synthesis in rat granulosa cells. Endocrinology 126, 2302–2306.PubMedCrossRefGoogle Scholar
  20. 20.
    Tse, W. T. and Forget, B. G. (1990) Reverse transcription and direct amplification of cellular RNA transcripts by Taq polymerase. Gene 88, 293–296.PubMedCrossRefGoogle Scholar
  21. 21.
    Rappolee, D. F. A., Mark, D., Banda, M. I., and Werb, Z. (1988) Wound macro-phages express TGF-± and other growth factors in vivo: analysis by mRNA phenotyping. Science 241, 708–712.PubMedCrossRefGoogle Scholar
  22. 22.
    Rauno, G., Brash, D. E., and Kidd, K. K. (1991) PCR: the first few cycles. Perkin-Elmer Cetus Amplifications 7, 1–4.Google Scholar
  23. 23.
    Grouse, J. and Amorese, D. (1987) Ethanol precipitation: ammonium acetate as an alternative to sodium acetate. BRL Focus 9, 3–5.Google Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1996

Authors and Affiliations

  • Beverly C. Delidow
    • 1
  • John P. Lynch
    • 2
  • John J. Peluso
    • 2
  • Bruce A. White
    • 2
  1. 1.Department of AnatomyUniversity of Connecticut Health CenterFarmingtonUK
  2. 2.Department of AnatomyUniversity of Connecticut Health CenterFarmington

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