Department of Biotechnology, Indian Institute of Technology Madras, Chennai, India.
Nanotechnology. 2010 Jun 25;21(25):255704. doi: 10.1088/0957-4484/21/25/255704. Epub 2010 Jun 2.
Improvement of the specificity and efficiency of the polymerase chain reaction (PCR) by nanoparticles is an emerging area of research. We observed that TiO(2) nanoparticles of approximately 25 nm diameter caused significant enhancement of PCR efficiency for various types of templates (namely plasmid DNA, genomic DNA and complementary DNA). By a series of experiments, the optimal TiO(2) concentration was determined to be 0.4 nM, which resulted in up to a seven-fold increase in the amount of PCR product. As much as 50% reduction in overall reaction time (by reduction of the number of cycles and the time periods of cycles) was also achieved by utilizing TiO(2) nanoparticles without compromising the PCR yield. Investigations of the mechanism of such PCR enhancement by simulations using the 'Fluent K epsilon turbulent model' provided evidence of faster heat transfer in the presence of TiO(2) nanoparticles. Consistent with these findings, TiO(2) nanoparticles were observed to augment the denaturation of genomic DNA, indicating more efficient thermal conductivity through the reaction buffer. TiO(2) nanoparticle-assisted PCR may be useful for profound reduction of the overall PCR reaction period and for enhanced amplification of DNA amplicons from a variety of samples, including GC-rich templates that are often observed to yield unsatisfactory results.
通过纳米粒子提高聚合酶链反应（PCR）的特异性和效率是一个新兴的研究领域。我们观察到，直径约为 25nm 的 TiO(2)纳米粒子显著提高了各种类型模板（即质粒 DNA、基因组 DNA 和互补 DNA）的 PCR 效率。通过一系列实验，确定了最佳的 TiO(2)浓度为 0.4nM，这使得 PCR 产物的量增加了 7 倍。通过使用 TiO(2)纳米粒子，还可以将总反应时间（通过减少循环次数和循环时间来实现）缩短多达 50%，而不会影响 PCR 产量。利用“Fluent K epsilon 湍流模型”进行模拟的研究结果表明，TiO(2)纳米粒子的存在可以实现更快的热传递。这些发现表明，TiO(2)纳米粒子可以增强基因组 DNA 的变性，这表明通过反应缓冲液的热导率更高。TiO(2)纳米粒子辅助 PCR 可能有助于大大缩短整个 PCR 反应周期，并增强从各种样品（包括富含 GC 的模板）中扩增 DNA 扩增子，这些模板通常会产生不理想的结果。
