Cao Xueyan, Shi Xiangyang, Yang Wenchao, Zhang Xiaodong, Fan Chunhai, Hu Jun
Nanobiology Laboratory, Bio-X Life Science Research Center, College of Life Science and Biotechnology, Shanghai JiaoTong University, Shanghai, 200240, PR China.
Analyst. 2009 Jan;134(1):87-92. doi: 10.1039/b812176a. Epub 2008 Oct 18.
The polymerase chain reaction (PCR) has become a fundamental technique in molecular biology. Improvements to enhance the specificity and efficiency of PCR are always required and remain a great challenge. Here we introduce generation 4 (G4) and 5 (G5) poly(amidoamine) (PAMAM) dendrimers and their derivatives with different terminal groups as a novel class of enhancers to improve the specificity and efficiency of PCR. We show that the dendrimers are effective in optimizing error-prone two-round PCR and non-specific PCR in specificity and efficiency. With the increase of the number and density of terminal amine groups, the optimum concentration of polycationic dendrimers could be decreased to as low as 1.35 nM, which is four orders of magnitude smaller than that of fully acetylated dendrimers and carboxyl-terminated dendrimers. The present study underlines a fact that dendrimers could be used as a powerful additive to enhance the specificity and efficiency of PCR.
聚合酶链反应（PCR）已成为分子生物学中的一项基础技术。始终需要对PCR进行改进以提高其特异性和效率，而这仍然是一项巨大的挑战。在此，我们引入第4代（G4）和第5代（G5）聚（酰胺胺）（PAMAM）树枝状大分子及其具有不同端基的衍生物，作为一类新型增强剂，以提高PCR的特异性和效率。我们表明，树枝状大分子在优化易错两轮PCR和非特异性PCR的特异性和效率方面是有效的。随着末端胺基数量和密度的增加，聚阳离子树枝状大分子的最佳浓度可降至低至1.35 nM，这比完全乙酰化树枝状大分子和羧基末端树枝状大分子的最佳浓度小四个数量级。本研究强调了一个事实，即树枝状大分子可用作增强PCR特异性和效率的强大添加剂。
