聚合酶链式反应热流变化的DSC实验研究

在PCR每个循环中,目的基因在DNA聚合酶的催化作用下实现快速扩增,同时伴随着化学键的断裂和生成,而不同循环数的扩增效率不同,引起的热现象也不同。实验通过差示扫描量技术,以HBV为PCR扩增体系,分别研究了变性、退火和延伸阶段的热焓及其随循环数的变化,通过分析得出：变性阶段是放热过程,第17个循环放热量达到最大,退火和延伸阶段是吸热过程;3个阶段的热焓随循环数增加都发生明显的变化,其中变性阶段的热流变化最关键。

Research for the variation of enthalpy in PCR with DSC

The target gene rapid amplification is always accompanied by the rupture and production of chemical bond in entire PCR process,the thermal phenomenon is changed with variable amplification efficiency in different time.The HBV as the target gene in PCR was replicated.The variation of enthalpy in denaturation,annealing and extension phase were studied by DSC.The result of analysis showed that the denaturation phase of the reaction was endothermic process.When the cycle number was 17,the yield of emission reached the peak,and the annealing and extension phase was heat emission process.The enthalpy changed differently with the cycle number increase in these phase.It indicated the denaturation was crucial.