Ma Lu, He Shibin, Huang Jing, Cao Lei, Yang Fei, Li Lijia
Wuhan University, People's Republic of China.
Biochimie. 2009 Aug;91(8):969-73. doi: 10.1016/j.biochi.2009.04.020. Epub 2009 May 13.
We found that semiconductor quantum dots (QDs) dramatically improved both product yield and specificity of PCR. The concentration of QDs is important for improving PCR amplification. In the presence of appropriate concentration of mercaptoacetic acid (MAA)-coated QDs, specificity and yield of PCR were enhanced. Also, strong nonspecific bands and weaker smeared bands were eliminated. At lower annealing temperatures (25-45 degrees C), addition of MAA-coated QDs into the PCR reagent produced specific PCR products without nonspecific sequence amplification. MAA alone did not improve PCR amplification. Streptavidin (SA) surface modified QDs with different size also effectively improved the specificity of PCR, demonstrating that the observed effect was not due to property of the QD surface but instead due to the QD itself. Bovine Serum Albumin (BSA) could relieve Taq polymerase from MAA-coated QDs in PCR by interaction with QDs and therefore imply that QDs improve specificity of PCR by interaction with Taq polymerase. These results demonstrate that QDs, added to reaction mixes at appropriate concentrations, can increase PCR yield and improve PCR specificity, even at low annealing temperatures. We assume that many different surface modified polymeric nanoparticles might have similar effects.
我们发现半导体量子点(QDs)显著提高了PCR的产物产量和特异性。量子点的浓度对于改善PCR扩增很重要。在存在适当浓度的巯基乙酸(MAA)包被的量子点的情况下,PCR的特异性和产量得到了提高。此外,强非特异性条带和较弱的拖尾条带被消除。在较低的退火温度(25 - 45摄氏度)下,向PCR试剂中添加MAA包被的量子点可产生特异性PCR产物,而无非特异性序列扩增。单独的MAA并不能改善PCR扩增。用不同大小的链霉亲和素(SA)表面修饰的量子点也有效地提高了PCR的特异性,这表明观察到的效果不是由于量子点表面的性质,而是由于量子点本身。牛血清白蛋白(BSA)可通过与量子点相互作用,在PCR中使Taq聚合酶从MAA包被的量子点上解离,因此意味着量子点通过与Taq聚合酶相互作用提高了PCR的特异性。这些结果表明,以适当浓度添加到反应混合物中的量子点可以提高PCR产量并改善PCR特异性,即使在低退火温度下也是如此。我们推测许多不同表面修饰的聚合物纳米颗粒可能具有类似的效果。