Effects of different concentrations and sizes of UCNPs on PCR amplification specificity. Three commercial real-time PCR master mixes were applied to determine the effects of different concentrations and sizes of UCNPs on the specificity of PCR amplification of a 120 bp 5S rDNA fragment from soybean genomic <t>DNA,</t> at an annealing temperature of 50°C. ( A ) AccuPower PCR PreMix with Top Polymerase (Bioneer). ( B ) AmpliTaq Gold 360 Master Mix with AmpliTaq Gold DNA Polymerase (Applied Biosystems). ( C ) <t>HotStarTaq</t> Plus Master Mix with HotStarTaq DNA Polymerase (Qiagen). Lane M: DNA marker; lane C: template without UCNPs; lanes 1∼5∶5, 7.5, 10, 15, and 30 nM of QDs, respectively; lanes 6∼8∶0.5, 0.75, and 1.0 µg/µL of 40-nm-sized UCNPs, respectively; lanes 9∼14∶1×( = 2.4×10 5 particles/µL), 10×, 15×, 20×, 25×, and 30× of 70-nm-sized UCNPs, respectively; lanes 15∼20∶1×, 10×, 15×, 20×, 25×, and 30× of 250-nm-sized UCNPs, respectively. 