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Polymerase Chain Reaction

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Molecular Biomethods Handbook

Part of the book series: Springer Protocols Handbooks ((SPH))

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Abstract

In general, advances in biological and biochemical research are brought about by tmprovements and refinement of methods in current use. There are times, however, when a technique is developed that revolutionizes a particular field of research The polymerase chain reaction, or PCR, developed at the Cetus Corporation in 1985 is one such example The technique enables large amounts of DNA to be produced from very small amounts of starting material and mimics the basic mechamsm of DNA replication and the manner in which it is carried out. The PCR was first described in 1985 and is a technique resulting in near exponential enzymatic amplification of DNA to a level easily detected by conventional methods, such as gel electrophoresis (2).

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References

  1. Saiki, R. K., Scharf, S. J., Faloona, F., Mullis, K. B., Horn, G. T., Erlich, H. A., and Amheim, N. (1985) Enzymatic amplification of 8 globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 230, 1350–1354.

    Article  PubMed  CAS  Google Scholar 

  2. Paabo, S. (1989) Ancient DNA extraction characterisation molecular cloning and amplification. Proc. Natl. Acad. Sci. USA 86, 1939–1943.

    Article  PubMed  CAS  Google Scholar 

  3. Rychlik, W. (1994) New algorithm for determining primer efficiency in PCR and sequencing. J. NIH Res. 6,78.

    Google Scholar 

  4. Knoth, K., Roberds, S., Poteet, C., and Tamkun, M. (1988) Highly degenerate inosine containing primers specifically amplify rare cDNA using the polymerase chain reaction.Nucleic Acids Res. 16,932.

    Article  Google Scholar 

  5. Reischl, U. and Kochanowski, B. (1995) Quantitative PCR A survey of the present technology. Mol. Biotechnol. 3,55–71.

    Article  PubMed  CAS  Google Scholar 

  6. Eckert, K. A. and Kunkel, T. A. (1990) High fidelity DNA synthesis by the Thermus aquaticus DNA polymerase. Nucleic Acids Res. l8 3739–3744.

    Article  Google Scholar 

  7. Myers, T. W. and Gelfand, D. H. (1991) Biochemistry 30,7661.

    Article  PubMed  CAS  Google Scholar 

  8. Cheng, S., Cheng, S-Y., Gravitt, P., and Respess, R. (1994) Long PCR. Nature 369, 684–685.

    Article  PubMed  CAS  Google Scholar 

  9. Hung, T, Mak, K, and Fong, K (1990) A speclficlty enhancer for the polymerase cham reactlon Nucleic Aczds Res 18, 1666

    Article  Google Scholar 

  10. Chou, Q, Russell, M, Birch, D E, Raymond, J and Bloch, W (1992) Prevention of pre-PCR mlsprlming and primer dlmerlzatlon improves low copy number amphficatlon Nuclezc Aczds Res 20, 1717–1723

    Article  CAS  Google Scholar 

  11. Chamberlam, J S, Gibbs, R A, Ranier, J E, Nguyen, P N, and Caskey, C T (1988)Deletion screening of the Duchenne muscular dystrophy locus via multiplex DNA ampltficatlon Nuclerc Acids Res 16, 11,141-l 1,156

    Google Scholar 

  12. Marmo, J, yatt, P, Flood, S J, and McBride, L (1997) A TaqMan multiplex PCR mesenger RNA assay Clzn Chem

    Google Scholar 

  13. Kwok, S and Hlguchl, R (1989) Avoiding false posltlves with PCR Nature 339,237–238

    Article  PubMed  CAS  Google Scholar 

  14. Longo, N, Bermnger, N S, and Hartley, J L (1990) Use of uracll N-glycolyase to control carry-over contammatlon in polymerase chain reaction Gene 93, 125–128

    Article  PubMed  CAS  Google Scholar 

  15. Kawasaki, E S (1990) Amphficatlon of RNA, in PCR Protocols A Guide to Methods & Applrcatzons (Inms, A, Gelfand, D, Smnsky, J J, and White, T J, eds ), Academic, San Diego, CA, pp 21–27

    Google Scholar 

  16. Liang, P and Pardee, A B (1992) Differenyial display of eukaryotic messenger RNA by means of the polymerase cham reaction Science 257, 967–971

    Article  PubMed  CAS  Google Scholar 

  17. Bevan, I S, Rapley, R, and Walker, M R (1992) Sequencing of PCR-amplified DNA PCR Methods Appl 1 (4), 222–227

    CAS  Google Scholar 

  18. Gyllenstem, U B and Erhch, H A (1988) Generation of single stranded DNA by the polymerase chain reaction and Its apphcatlon to direct sequencing of the HLA-DQA locus Proc Natl Acad Scr USA 85,7652–7656

    Article  Google Scholar 

  19. McConlogue, L, Brow, M A D, and Inms, M A (1988) Structure-mdependent DNA amplification by PCR using 7-deaza-2′deoxyguanosme Nucleic Acids Res 16, 9869

    Article  PubMed  CAS  Google Scholar 

  20. Rapley, R (1996) Methods zn Molecular Bzology, vol 65 PCR Sequenczng Protocols Humana, Totowa, NJ

    Google Scholar 

  21. Slatko, B E (1996) Thermal cycle dldeoxy DNA sequencing MoZ Bzotech 6,311–322

    Article  CAS  Google Scholar 

  22. Mitchell, L G and Meml, C R (1989) Affinity generation of single stranded DNA for dldeoxy sequencing following the polymerase chain reaction Anal Bzochem 178,239–242

    Article  CAS  Google Scholar 

  23. Stoflet, E S, Koeberl, D D, Sarker, G, and Sommer, S S (1988) Genomlc amphficatlon with transcript sequencing Science 239,491–494

    Article  PubMed  CAS  Google Scholar 

  24. White, B A, ed (1996) Methods zn Molecular Bzology vol 67 PCR Clonzng Protocols From Molecular Clonrng to Genetic Engzneerzng Humana, Totowa, NJ

    Google Scholar 

  25. Mead, D A, Pey, N K, Herrnstadt, C, Marc& R A, and Smith, L in (1991) A umversal method for the direct cloning of PCR amplified nucleic acid Bzo/Technology 9, 657–663

    Article  CAS  Google Scholar 

  26. Hlguchl, R, Krummel, B, and Salki, R K (1988) A general method of in vitro preparatton and specific mutagenesis of DNA fragments study of protem and DNA mteractlons Nucleic Acids Res 16, 7351–7367

    Article  Google Scholar 

  27. Sarker, G and Sommer, S S (1990) The megaprlmer method of site directed mutagenesls Bzotechnzques 8,404–407

    Google Scholar 

  28. Clackson, T and Winter, G (1989) Sticky feet directed mutagenesis and its appllcatlons to swapping antibody domains Nucleic Aczds Res 17, 10,163-10,170

    Google Scholar 

  29. Cotton, R G H (1992) Detection of mutations in DNA Current Opzn Bzotechnol 3,24–30

    Article  CAS  Google Scholar 

  30. Newton, C R, Graham, A, Heptmstall, L E, Powell, S J, Summers, C, Kalsheker, N, Smith, J C, and Markham, A F (1989) Analysis of any point mutation in DNA The amplification refractory mutation system Nucleic Aczds Res 17,2503–2516

    Article  CAS  Google Scholar 

  31. Wartell, R in, Hosseml, S H, and Moran, C P Jr (1990) Detecting base pair substltutlons in DNA fragments by temperature gradlent gel electrophoresls Nuclezc Acrds Res 18, 2699–2705

    Article  CAS  Google Scholar 

  32. Dockhorn-Dwormczak, B, Dwormczak, B, Brommelkamp, L, Bulles, J, Horst, J, and Backer, W (1991) Non-lsotoplc detection of single strand conformatlonal polymorphism (PCR-SSCP) a rapid and sensltlve technique in the diagnosis of phenylketonurla Nuclei Aczds Res 19,2500–2505

    Article  Google Scholar 

  33. Ochman, H, Gerber, S A, and Hartl, D L (1988) Genetic apphcatlons of an inverse polymerase chain reaction Genetzcs 120,621–625

    CAS  Google Scholar 

  34. Gosden, J, ed (1997) Methods zn Molecular Biology, vol 71 PRINS and In Sztu PCR Protocols Humana, Totowa, NJ

    Google Scholar 

  35. Wmn-Deen, E S (1997) Automation of molecular genetic methods DNA ampllficatlon techniques J Urn Lzgand Assay 19,21–26

    Google Scholar 

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Authors and Affiliations

  1. University of Hertfordshzre, Hatfield, UK

    Ralph Rapley

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  1. Ralph Rapley
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Editor information

Editors and Affiliations

  1. University of Herfordshire, Hatfield, UK

    Ralph Rapley  & John M. Walker  & 

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© 1998 Humana Press Inc , Totowa, NJ.

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Rapley, R. (1998). Polymerase Chain Reaction. In: Rapley, R., Walker, J.M. (eds) Molecular Biomethods Handbook. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1007/978-1-59259-642-3_25

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  • DOI: https://doi.org/10.1007/978-1-59259-642-3_25

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-501-0

  • Online ISBN: 978-1-59259-642-3

  • eBook Packages: Springer Book Archive

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