单壁碳纳米管对长片段聚合酶链式反应(PCR)增效机理的初步研究

摘要

为了开发新型基因扩增技术,采用两步PCR (two-step PCR)和rpsL保真度实验等方法,对单壁碳纳米管(single-wall carbon nanotube,SWCNT)在长片段PCR反应中的增效机理进行研究.结果表明:在长片段PCR反应中,单壁碳纳米管最适工作质量浓度为0.8 mg/mL;单壁碳纳米管可以在较短的延伸时间增加扩增产物产量,提高DNA聚合酶的反应效率；通过rpsL保真度实验显示,对照组PCR产物的错误率为5.32 × 10-6,而实验组的错误率仅为2.39×10-6.%The effect of single-wall carbon nanotube (SWCNT) on long-PCR was studied with a two-step PCR and rpsL fidelity assay. The experiments results showed that the best concentration of SWCNT was 0.8 mg/mL in long-PCR system. SWCNT did contribute to the high activity of DNA polymerase, so an amplified DNA product was observed even within a very short reaction time. The error frequency of PCR products was 5.32 × 10-6 in the control group, while the error frequency declined to 2.39 × 10-6 by using SWCNT.