Maximizing specificity and yield of PCR by the quantum dot itself rather than property of the quantum dot surface.
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.
Ma
Lu
L
Wuhan University, People's Republic of China.
He
Shibin
S
Huang
Jing
J
Cao
Lei
L
Yang
Fei
F
Li
Lijia
L
D000818:Animals; D002417:Cattle; D010316:Particle Size; D016133:Polymerase Chain Reaction / Q000379:methods*; D011108:Polymers / Q000737:chemistry; D045663:Quantum Dots; D012710:Serum Albumin, Bovine / Q000378:metabolism; D013499:Surface Properties; D019914:Taq Polymerase / Q000378:metabolism; D013696:Temperature; D013864:Thioglycolates / Q000737:chemistry
D011108:Polymers; D013864:Thioglycolates; D012710:Serum Albumin, Bovine; C017487:2-mercaptoacetate; D019914:Taq Polymerase