Improved thermal cycling durability and PCR compatibility of polymer coated quantum dot.
Quantum dots have experienced rapid development in imaging, labeling and sensing in medicine and life science. To be suitable for polymerase chain reaction (PCR) assay, we have tested QD thermal cycling durability and compatibility, which have not been addressed in previous reports. In this study, we synthesized CdSe/ZnS QDs with a surface modification with high-MW amphiphilic copolymers and observed that Mg²⁺ ions in the PCR reaction could induce the QDs to precipitate and reduce their fluorescence signal significantly after thermal cycling. To overcome this problem, we used mPEG2000 to conjugate the QD surface for further protection, and found that this modification enables QDs to endure 40 thermal cycles in the presence of other components essential for PCR reactions. We have also identified that QDs have different effects on rTaq and Ex Taq polymerization systems. A high QD concentration could apparently reduce the PCR efficiency, but this inhibition was relieved significantly in the Ex PCR system as the concentration of Ex Taq polymerase was increased. Real-time PCR amplification results showed that QDs could provide a sufficiently measurable fluorescence signal without excessively inhibiting the DNA amplification. Based on this improved thermal cycling durability and compatibility with the PCR system, QDs have the potential to be developed as stable fluorescent sensors in PCR and real-time PCR amplification.
Key Laboratory of Medical Cell Biology-Ministry of Education, Department of Biochemistry and Molecular Biology, China Medical University, Shenyang 110001, People's Republic of China.
Zhe
Z
Xun
Xiaoyun
X
Zhao
Yifu
Y
Guan
D019187:Cadmium Compounds / Q000737:chemistry; D005453:Fluorescence; D011092:Polyethylene Glycols / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000379:methods*; D011108:Polymers / Q000737:chemistry*; D045663:Quantum Dots; D018036:Selenium Compounds / Q000737:chemistry; D013440:Sulfides / Q000737:chemistry; D013696:Temperature; D017967:Zinc Compounds / Q000737:chemistry
D019187:Cadmium Compounds; D011108:Polymers; D018036:Selenium Compounds; D013440:Sulfides; D017967:Zinc Compounds; D011092:Polyethylene Glycols; C028210:monomethoxypolyethylene glycol; C058667:cadmium selenide; C031238:zinc sulfide