A hot start alternative for high-fidelity DNA polymerase amplification mediated by quantum dots  

作　　者：Fuming Sang Yang Yang Ying Lin Zhizhou Zhang 

机构地区：[1]School of Marine Science and Technology, Harbin Institute of Technology, Weihai 264209, China [2]Institute of Biological Sciences and Biotechnology, College of Chemistry, Chemical Engineering and Biotechnology, Donghua University,Shanghai 201620, China

出　　处：《Acta Biochimica et Biophysica Sinica》2014年第6期502-511,共10页生物化学与生物物理学报（英文版）

基　　金：This work was supported by the grants from the National Natural Science Foundation of China （No.31071170）, GRED BIO fund, Harbin Institute of Technology （WH） （IMJQ 10070012）, Harbin Institute of Technology （hitwh200904）, 985 Fund, Harbin Institute of Technology-NSRIF （2011101）, and The Science and Technology Development Projects of Weihai （2011DXGJ 13, 2012DXGJ02）.

摘　　要：Quantum dots （QDs） are of great interest due to their unique chemical and physical properties. Recently, a hot start （HS） polymerase chain reaction （PCR） amplification performance based on QDs with a high-fidelity Pfu DNA polymerase has been reported. However, whether QDs can trigger HS effects with other high-fidelity or conventional DNA polymerases is yet to be understood. In the present study, we studied the QD-triggered HS effects with four high-fidelity and three conventional DNA polymerases, and the HS effect comparisons among them were also made. It was found that QDs could trigger a distinct HS PCR amplification performance with all the four tested high,fidelity DNA polymerases, and specific target DNA could be well amplified even if the PCR mixture was preincubated for 2 h at 50℃. On the contrary, the HS effects were not prominent with all the three conventional Taq DNA polymerases. Specifically, the fidelity of Pfu is not sacrificed in the presence of QDs, even after a 1 h pre-incu- bation at 50℃ before PCR. Furthermore, the electrophoresis results preliminarily demonstrated that QDs prefer to adsorb high-fidelity polymerases rather than conventional ones, which might result in the QD-triggered HS effects on PCR performance by using high-fidelity DNA poly- merases.Quantum dots （QDs） are of great interest due to their unique chemical and physical properties. Recently, a hot start （HS） polymerase chain reaction （PCR） amplification performance based on QDs with a high-fidelity Pfu DNA polymerase has been reported. However, whether QDs can trigger HS effects with other high-fidelity or conventional DNA polymerases is yet to be understood. In the present study, we studied the QD-triggered HS effects with four high-fidelity and three conventional DNA polymerases, and the HS effect comparisons among them were also made. It was found that QDs could trigger a distinct HS PCR amplification performance with all the four tested high,fidelity DNA polymerases, and specific target DNA could be well amplified even if the PCR mixture was preincubated for 2 h at 50℃. On the contrary, the HS effects were not prominent with all the three conventional Taq DNA polymerases. Specifically, the fidelity of Pfu is not sacrificed in the presence of QDs, even after a 1 h pre-incu- bation at 50℃ before PCR. Furthermore, the electrophoresis results preliminarily demonstrated that QDs prefer to adsorb high-fidelity polymerases rather than conventional ones, which might result in the QD-triggered HS effects on PCR performance by using high-fidelity DNA poly- merases.

关 键 词：quantum dot hot start PCR high-fidelityDNA polymerase TAQ 

分 类 号：Q754[生物学—分子生物学] O471.1[理学—半导体物理]