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The Effect of Temperature and Oligonucleotide Primer Length on the Specificity and Efficiency of Amplification by the Polymerase Chain Reaction
Abstract
The polymerase chain reaction (PCR) is most effectively performed using a thermostable DNA polymerase such as that isolated from Thermus aquaticus. Since temperature and oligonucleotide length are known to control the specificity of oligonucleotide hybridization, we have investigated the effect of oligonucleotide length, base composition, and the annealing temperature on the specificity and efficiency of amplification by the PCR. Generally, the specificity of PCR is controlled by the length of the oligonucleotide and/or the temperature of annealing of the primer to the template. An empirical relationship between oligonucleotide length and ability to support amplification was determined. This relationship allows for the design of specific oligonucleotide primers. A model is proposed which helps explain the observed dependence of PCR on annealing temperature and length of the primer.
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Cites
Primer-Directed Enzymatic Amplification of DNA with a Thermostable DNA Polymerase
Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. J., Higuchi, R., Horn, G. T., Mullis, K. B., & Erlich, H. A. (1988) · Science · doi:10.1126/science.2448875 · Cited by 17,719
Saiki, R. K., Scharf, S., Faloona, F., Mullis, K. B., Horn, G. T., Erlich, H. A., & Arnheim, N. (1985) · Science · doi:10.1126/science.2999980 · Cited by 8,956
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Cited by
Multiplex PCR: Optimization and Application in Diagnostic Virology
Elnifro, E. M., Ashshi, A. M., Cooper, R. J., & Klapper, P. E. (2000) · Clinical Microbiology Reviews · doi:10.1128/cmr.13.4.559-570.2000 · Cited by 964
Metadata
- Authors
- WU, D. Y., UGOZZOLI, L., PAL, B. K., QIAN, J., & WALLACE, R. B.
- Venue
- DNA and Cell Biology
- Publisher
- SAGE Publications
- Published
- 1991-04
- Volume
- 10
- Issue
- 3
- Pages
- 233-238
- ISSN
- 1044-5498 (ISSN-L)
- 1557-7430
- OpenAlex
- W2109167494
- Cited by
- 192
- References
- 2
- FWCI
- 2.28
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.liebertpub.com/nv/resources-tools/text-and-data-mining-policy/121/ (tdm)
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- WU, D. Y., UGOZZOLI, L., PAL, B. K., QIAN, J., & WALLACE, R. B. (1991). The Effect of Temperature and Oligonucleotide Primer Length on the Specificity and Efficiency of Amplification by the Polymerase Chain Reaction. DNA and Cell Biology. https://doi.org/10.1089/dna.1991.10.233
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Bibliographic data: Semantic Scholar, OpenAlex, Crossref.
Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
