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The effect of temperature and oligonucleotide primer length on the specificity and efficiency of amplification by the polymerase chain reaction

plr0201 添加于 2015-4-28 18:27 | 1198 次阅读 | 0 个评论

作者
Wu DY, Ugozzoli L, Pal BK, Qian J, Wallace RB
摘要
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.
详细资料
- 关键词: Base Sequence; DNA/genetics/isolation & purification; DNA-Directed DNA Polymerase; *Gene Amplification; Humans; Molecular Sequence Data; *Oligonucleotide Probes; Polymerase Chain Reaction; Substrate Specificity; Taq Polymerase; Temperature
- 文献种类:期刊
- 期刊名称: DNA and Cell Biology
- 期刊缩写: DNA Cell Biol
- 期卷页: 1991年第10卷第3期 233-238页
- 地址: Department of Molecular Biochemistry, Beckman Research Institute of the City of Hope, Duarte, CA 91010
- ISBN: 1044-5498
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