摘要
“双退火温度”多聚酶链反应扩增丙型肝炎病毒ＮＳ_５基因李庆生，郭建平，陶其敏（北京医科大学人民医院肝病研究所，北京１０００４４）我们在扩增丙型肝炎病毒ＮＳ５基因中，探讨了退火温度与扩增产物特异性之间的关系，发现“双退火温度”多聚酶链反应可很好地解决目?..
The polymerase chain reaction is a very powerful technique in molecular biology. It has greatly simpified the molecular clone protocol. But sometimes, the introduction of restriction endonuclase sites into the primers、the high mutation of target gene、larger product of PCR and shorter primers,etc. can lead to the failure of amplification. During the amplification of Hepatitis C Virus NS_5 gene fragment,we discovered that“two annealing temperature PCR”. which at first amplified the target gene at lower annealing temperature so as to increase the copy of target gene,then enhanced the annealing temperature to guarantee the specificity of amphfication,was a practical and useful method,especially for unideal primers.
关键词
丙型肝炎病毒
NS5基因
聚合酶链反应
Two annealing temperature,Polymerase chain reaction,NS_5 gene,Hepatitis C virus
