Development and Application of Real-Time Quantitative PCR

  • P. Mickey Williams
  • Todd Giles
  • Ayly Tucker
  • Jane Winer
  • Chris Heid
Part of the Advanced Biomedical Technologies book series (ABT)

Abstract

Since the original description of the polymerase chain reaction (PCR), novel applications of this technique have been reported at an exponential rate. The power of the method lies in the ability to analyze minute amounts of samples (e. g., dried blood, microliters of liquid, several cells) or even to detect a single molecule. Virtually every area of biological science has been impacted by PCR, including forensics, genetics, clinical diagnostics, drug discovery, and environmental biology.

Keywords

Obesity Phenol Leukemia Recombination Titration 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Bassler HA, Flood SJ, Livak KJ, Marmaro J, Knorr R, and Batt CA (1995): Use of a fluorogenic probe in a PCR-based assay for the detection of Listeria monocytogenes. Appl Environ Microbiol 61:3724–8.PubMedGoogle Scholar
  2. Becker-Andre M (1989): Absolute mRNA quantification using the polymerase chain reaction (PCR). A novel approach by a PCR aided transcript titration assay (PATTY). Nucleic Acids Res 17:9437–46.PubMedCrossRefGoogle Scholar
  3. Becker-Andre M (1991): Quantitative evaluation of mRNA levels. Meth Mol Cell Biol 2:189–201.Google Scholar
  4. Becker-Andre M (1993): Absolute levels of mRNA by polymerase chain reaction-aided transcript titration assay. Meth Enzymol 218:420–45.PubMedCrossRefGoogle Scholar
  5. Ferré F (1992): Quantitative or semi-quantitative PCR: reality versus myth [Review]. PCR Meth Appl 2:1–9.Google Scholar
  6. Gibson UEM, Heid C, and Williams PM (1996): A novel method for real time quantitative RT-PCR. Genome Res 6:995–1001.PubMedCrossRefGoogle Scholar
  7. Heid C, Stevens J, Livak K, and Williams PM (1996): Real time quantitative PCR. Genome Res 6:986–94.PubMedCrossRefGoogle Scholar
  8. Higuchi R, Dollinger G, Walsh PS, and Griffith R (1992): Simultaneous amplification and detection of specific DNA sequences. Biotechnology 10:413–17.PubMedCrossRefGoogle Scholar
  9. Holland PM, Abramson RD, Watson R, and Gelfand DH (1991): Detection of specific polymerase chain reaction product by utilizing the 5´-3´ exonuclease activity of Thermus aquaticus DNA polymerase. Proc Natl Acad Sci USA 88:7276–80.PubMedCrossRefGoogle Scholar
  10. Kellogg DE, Sninsky JJ, and Kwok S (1990): Quantitation of HIV-1 proviral DNA relative to cellular DNA by the polymerase chain reaction. Anal Biochem 189:202–8.PubMedCrossRefGoogle Scholar
  11. King KL, Jadine L, Winer J, Luis E, Yen R, Hooley J, Williams PM, and Mather J (1996): Cardiac fibroblasts produce leukemia inhibitory factor and endothelin, which combine to induce cardiac myocyte hyperthrophy in vitro. Endocrine 5:85–93.PubMedCrossRefGoogle Scholar
  12. Lee LG, Connell CR, and Bloch W (1993): Allelic discrimination by nick-translation PCR with fluorogenic probes. Nucleic Acids Res 21: 3761–6.PubMedCrossRefGoogle Scholar
  13. Livak KJ, Flood SJ, Marmaro J, Giusti W, and Deetz K (1995a): Oligonucleotides with fluorescent dyes at opposite ends provide a quenched probe system useful for detecting PCR product and nucleic acid hybridization. PCR Meth Appl 4:357–62.Google Scholar
  14. Livak KJ, Marmaro J, and Todd JA (1995b): Towards fully automated genome-wide polymorphism screening [letter]. Nat Genet 9:341–2.PubMedCrossRefGoogle Scholar
  15. Luoh S-M, Di Marco F, Levin N, Armanini M, Xie M-H, Armanini M, Xie M-H, Nelson C, Bennett GL, Williams PM, Spencer SA, Gurney A, and de Sauvage FJ (1997): Cloning and characterization of a human leptin receptor using a biologically active leptin immunoadhesin. J Mol Endocrinol 18:77–85.PubMedCrossRefGoogle Scholar
  16. Pang S, Koyanagi Y, Miles S, Wiley C, Vinters HV, and Chen IS (1990): High levels of unintegrated HIV-1 DNA in brain tissue of AIDS dementia patients. Nature 343:85–9.PubMedCrossRefGoogle Scholar
  17. Piatak MJ, Luk KC, Williams B, and Lifson JD (1993): Quantitative competitive polymerase chain reaction for accurate quantitation of HIV DNA and RNA species. Biotechniques 14:70–81.PubMedGoogle Scholar
  18. Raeymaekers L (1995): A commentary on the practical applications of competitive PCR. Genome Res 5:91–4.PubMedCrossRefGoogle Scholar

Copyright information

© Birkhäuser Boston 1998

Authors and Affiliations

  • P. Mickey Williams
    • 1
  • Todd Giles
    • 1
  • Ayly Tucker
    • 1
  • Jane Winer
    • 1
  • Chris Heid
    • 1
  1. 1.Genentech, Inc.South San FranciscoUSA

Personalised recommendations