水热
热启动PCR
聚合酶
多重位移放大
分子生物学
DNA聚合酶
聚合酶
底漆二聚体
聚合酶链反应
DNA
化学
抗体
生物
单克隆抗体
生物化学
DNA提取
套式聚合酶链反应
基因
遗传学
多重聚合酶链反应
作者
D.E. Kellogg,И. Н. Рыбалкин,S Chen,N. M. Mukhamedova,Т. Н. Власик,Paul D. Siebert,Alex Chenchik
出处
期刊:PubMed
日期:1994-06-01
卷期号:16 (6): 1134-7
被引量:120
摘要
The specificity and DNA yield of PCRs are often improved by the "hot start" technique and analogous methods. The intent of the approach is to eliminate or prevent the generation of nonspecific PCR templates that may be synthesized at ambient temperature prior to thermal cycling. Monoclonal antibodies (MAbs) raised in mice to the purified DNA polymerase of Thermus aquaticus (Taq) were selected for their ability to reversibly block polymerase activity. The MAbs, incubated with Taq DNA polymerase and added to PCR tubes at ambient temperature, yield specific DNA fragments upon amplification when using high numbers of temperature cycles and a very low copy number of target DNA in a complex DNA background. This approach, using the TaqStart Antibody, permits the preparation of reaction mixtures at ambient temperatures without the subsequent opening of reaction tubes, use of grease or waxes, or of degradative enzymes and deoxyribonucleotide analogs.
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