HBV C区基因扩增与PCR条件的优化

Genes of HBV C Region Amplifying and Optimization of PCR Conditions

目的:为提高PCR检测乙型肝炎病毒(HBV)的特异性和灵敏度,降低成本,对PCR条件进行优化.方法:将HBV特异性基因片段转化到大肠杆菌DH5α中,提取质粒,利用PCR扩增HBV C区基因,对PCR条件中的退火温度、Mg2+浓度进行优化,并比较3种不同Taq DNA聚合酶的灵敏度.结果:HBV C区基因扩增的最佳退火温度为58℃,最佳Mg2+浓度为1.5mmol/L,Biostar Taq DNA聚合酶扩增到10-6.讨论:对HBV C区基因进行了转化和PCR实验条件的优化,为扩大PCR在乙型肝炎病毒(HBV)检测领域中的应用提供实验依据.

To increase the speciality and sensitivity of PCR for the genes of HBV C region and to decrease the cost of PCR detecting HBV and to optimize the PCR conditions.Methods: HBV genes were transformed into E.coli,DH5α.Plasmid was extracted from those E.coli. And the genes of HBV C region were amplified.Then optimizing the temperature of annealing and Mg2+ density,and comparing the sensitivities of three kinds of different Taq DNA polymerase.The results showed that the optimal annealing temperature was 58℃,the o...