CdTe quantum dots enhance feasibility of EvaGreen-based real-time PCR with decent amplification fidelity

Mol Biotechnol. 2013 Jul;54(3):969-76. doi: 10.1007/s12033-013-9650-z.

Abstract

Quantitative real-time PCR (qPCR), as an important quantitative technique for nucleic acids, has been widely used in many fields including clinical diagnosis, molecular biology, and cancer research. However, non-specific amplification products are still a frequent problem in qPCR. In this study, we investigated the effects of QDs on real-time amplification based on either SYBR Green I or EvaGreen. It was found that QDs could raise the amplification sensitivity and thus enhance the efficiency using SYBR Green I detection system. In the case of EvaGreen detection systems, addition of QDs also led to a better correlation coefficient than without QDs. EvaGreen-based system gave sharper peaks for melting curves than SYBR Green I. The experiments indicated that the polymerase activity could be partially blocked by QDs at the pre-PCR temperatures, resulting in the improvement of PCR specificity. These results indicated that CdTe QDs could be used as a descent qPCR enhancer. Good amplification fidelity in QDs-facilitated qPCR was also a plus that has not been reported elsewhere.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Benzothiazoles
  • Cadmium Compounds / chemistry*
  • DNA / analysis
  • DNA / chemistry
  • DNA / genetics
  • Diamines
  • Electrophoresis, Agar Gel
  • Fluorescent Dyes / chemistry*
  • Humans
  • Linear Models
  • Organic Chemicals / chemistry
  • Quantum Dots*
  • Quinolines
  • Real-Time Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Sequence Alignment
  • Tellurium / chemistry*

Substances

  • Benzothiazoles
  • Cadmium Compounds
  • Diamines
  • Fluorescent Dyes
  • Organic Chemicals
  • Quinolines
  • SYBR Green I
  • DNA
  • Tellurium
  • cadmium telluride