不同分子量PVP对PCR扩增反应效率的影响

Study on the Effects of Polyvinylpyrrolidone in Different Molecular-weight on the Efficiency of Polymerase Chain Reaction

以人为设计的110bp单链DNA为模板,研究不同分子量聚乙烯基吡咯烷酮(Polyvinyl-pyrrolidone,PVP)对聚合酶链式反应(polymerase chain reaction,PCR)的影响。实验结果表明:0.5%的PVP K-30、PVP 8000、PVP 58000和PVP 630000均可显著提高PCR效率,并且这4种化合物提高PCR效率的能力,依次为PVP 630000>PVP 58000>PVP K-30和PVP 8000;然而,在相同浓度时,PVP K-40却显著抑制PCR扩增反应;另外,溶解曲线的实验结果显示,0.5%PVP 630000可显著降低PCR产物的Tm值,表明PVP 630000可通过与DNA相互作用,降低DNA结构稳定性,使得模板更易在PCR的变性阶段解链,最终提高PCR效率。上述研究结果表明,不同分子量的PVP对PCR扩增效率的影响不同,与其分子量大小密切相关。

The effect of different molecular-weight Polyvinylpyrrolidone（PVP） on PCR efficiency has been researched into by undertaking the 110 bp,single-stranded DNA amplification with quantitative real-time polymerase chain reaction（QPCR）.The results indicates that 0.5% PVP K-30,PVP 8000,PVP 58000 and PVP 630000 could effectively enhance the PCR efficiency,and the ability of different molecular-weight PVP to improve PCR efficiency is PVP 630000,PVP 58000,PVP K-30 and PVP 8000,respectively.However,PVP K-40 could significantly inhibit the generation of PCR product.In addition,the results of melting curve showed that 0.5% PVP 630000 could dramatically reduce the Tm value of PCR product.It suggestes that PVP 630000 could interact with DNA template to reduce the stability of DNA conformation,promote the unwinding of DNA template in the denaturation stage of PCR,then enhance the PCR efficiency.The results reveal that the effects of different PVP on the PCR efficiency is closely related to the molecular-weight.