Ali Zeeshan, Jin Gang, Hu Zhili, Wang Zhifei, Khan Muhammad Ammar, Dai Jianguo, Tang Yongjun
Postdoctoral Innovation Practice Base, Shenzhen Polytechnic, Shenzhen 518055, China.
Shenzhen Nanshan People's Hospital, Shenzhen 518052, China.
J Nanosci Nanotechnol. 2018 Dec 1;18(12):8029-8046. doi: 10.1166/jnn.2018.16390.
Polymerase Chain Reaction (PCR) is one of the most common technologies used in many laboratories to produce millions of copies of targeted nucleic acid under in vitro conditions. However, PCR faces multiple challenges including limited availability of DNA in the sample, high GC contents of the template, low efficiency, and specificity in amplification. Moreover, some DNA fragments are very difficult to amplify due to their secondary structure and high melting temperature requirement. To overcome these challenges, many approaches including the application of PCR additives in PCR mixture; change in instrument design; optimization of PCR system by using the accurate concentration of magnesium ions, primers, and cycle number; enzyme modification; and setting up the new touchdown and nested PCR strategies have been adopted. Although these approaches have enriched the output of PCR, they are not all-purpose and optimization can be case dependent. Nanometer-sized materials (nanomaterials) have offered a possible solution to these problems as these materials have exceptional physio-chemical properties as compared to macroscopic materials. Among these nanomaterials, silicon-based materials, carbon-based materials, semiconductor quantum dots (QDs), and some metals are well-known PCR enhancer. Hence, new PCR has been designed to utilize the unique properties of nanomaterial and is known as nanomaterial-assisted PCR or simply nanoPCR. Results of many studies have shown that the combination of these nanomaterials and biomolecules can mimic the DNA replication process successfully as present in the living organism. In this review, we have discussed the role of these different nanomaterials one by one and also discussed the mechanisms through which these nanomaterials enhance the efficiency of PCR.
聚合酶链反应(PCR)是许多实验室中最常用的技术之一,用于在体外条件下产生数百万份靶向核酸的拷贝。然而,PCR面临多重挑战,包括样品中DNA可用性有限、模板的高GC含量、扩增效率低和特异性差。此外,一些DNA片段由于其二级结构和高解链温度要求而很难扩增。为了克服这些挑战,人们采用了许多方法,包括在PCR混合物中应用PCR添加剂;改变仪器设计;通过使用准确浓度的镁离子、引物和循环数来优化PCR系统;酶修饰;以及建立新的降落PCR和巢式PCR策略。尽管这些方法丰富了PCR的产出,但它们并非万能,优化可能因情况而异。纳米材料因其与宏观材料相比具有特殊的物理化学性质,为这些问题提供了一种可能的解决方案。在这些纳米材料中,硅基材料、碳基材料、半导体量子点(QDs)和一些金属是众所周知的PCR增强剂。因此,人们设计了新的PCR来利用纳米材料的独特性质,即纳米材料辅助PCR或简称为纳米PCR。许多研究结果表明,这些纳米材料与生物分子的组合可以成功模拟活生物体中存在的DNA复制过程。在这篇综述中,我们逐一讨论了这些不同纳米材料的作用,并讨论了这些纳米材料提高PCR效率的机制。