THE EFFECT OF TEMPERATURE AND OLIGONUCLEOTIDE PRIMER LENGTH ON THE SPECIFICITY AND EFFICIENCY OF AMPLIFICATION BY THE POLYMERASE CHAIN-REACTION

被引：132

作者：

WU, DY ^{[1
]}

UGOZZOLI, L ^{[1
]}

PAL, BK ^{[1
]}

QIAN, J ^{[1
]}

WALLACE, RB ^{[1
]}

机构：

[1] CITY HOPE NATL MED CTR, BECKMAN RES INT, DEPT MOLEC BIOCHEM, 1450 E DUARTE RD, DUARTE, CA 91010 USA

来源：

DNA AND CELL BIOLOGY | 1991年 / 10卷 / 03期

关键词：

D O I：

10.1089/dna.1991.10.233

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

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.

引用

页码：233 / 238

页数：6

共 30 条

[1] THE HIGHLY POLYMORPHIC REGION NEAR THE HUMAN INSULIN GENE IS COMPOSED OF SIMPLE TANDEMLY REPEATING SEQUENCES [J].