氧化石墨烯对聚合酶链式反应扩增效率、灵敏度和特异性的影响

发布时间：2018-06-12 18:21

本文选题：氧化石墨烯 + 聚合酶链式反应　；参考：《环境化学》2017年04期

【摘要】：以人类基因组DNA和志贺氏菌粗制裂解菌液为模板,扩增115 bp的Alu基因片段和402 bp的ipaH基因片段.通过设置氧化石墨烯(Graphene Oxide,GO)和模板的浓度为变量,分别研究了氧化石墨烯对聚合酶链式反应(Polymerase chain reaction,PCR)扩增效率和灵敏度的影响;运用多轮PCR扩增方法研究了氧化石墨烯对聚合酶链式反应扩增特异性的影响.结果显示,氧化石墨烯有效提高PCR扩增效率的范围为9—14倍,提高PCR灵敏度一个数量级以及增加多轮PCR扩增中的两轮扩增反应.从原子力显微镜(Atomic force microscope,AFM)的结果可以看出,本文制备的氧化石墨烯片层厚度主要集中在0.5—0.8 nm,少数大于1 nm,总体具有较大的比表面积.除此之外,本文还发现当氧化石墨烯浓度到达μg·μL~(-1)后,引物二聚体与氧化石墨烯的浓度呈正比,说明氧化石墨烯优化PCR的主要影响因素是其与引物的相互作用.
[Abstract]:The 115bp Alu gene fragment and the 402bp IpaH gene fragment were amplified from human genomic DNA and crude lytic liquid of Shigella spp. By setting the concentration of graphene oxide and template as variables, the effects of graphene oxide on the amplification efficiency and sensitivity of Polymerase chain reactionn (PCRs) were studied. The effects of graphene oxide on the amplification specificity of polymerase chain reaction were studied by multiplex PCR. The results showed that graphene oxide could effectively increase the PCR amplification efficiency by 9-14 times, increase the PCR sensitivity by one order of magnitude and increase the two-round amplification reaction in multi-round PCR amplification. It can be seen from the results of atomic force microscope (AFM) that the thickness of graphene oxide is mainly 0.5-0.8 nm, a few larger than 1 nm, and the total surface area is larger. In addition, when the concentration of graphene oxide reached 渭 g 渭 L ~ (-1), it was found that the concentration of the primer dimer was proportional to the concentration of graphene oxide, indicating that the interaction between the primer and the primer was the main influencing factor of the optimization of graphene oxide PCR.
【作者单位】：成都中医药大学医学技术学院;
【基金】：国家自然科学基金(81503366) 四川省教育厅项目(15ZB0098) 高等学校博士学科点专项科研基金(20135132120013) 成都中医药大学大学生科研实践创新基金(ky2016-018)资助~~
【分类号】：R440;O613.71

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