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	Characterization of the 5 ' to 3 ' nuclease activity of Thermus aquaticus DNA polymerase on fluorogenic double-stranded probes
	Huang, Qiuying ; Li, Qingge ; Li QG(李庆阁)
	2009
关键词	Taq DNA polymerase 5 '-Nuclease Real-time PCR Double-stranded probes Displacing probes MALDI-TOF MS
英文摘要	Taq DNA polymerase contains a polymerase domain for synthesizing new DNA strands and a 5'-nuclease domain for cleaving damaged DNA strands or RNA primers. Both of these domains play key roles in nucleic acid amplification and detection, especially in fluorogenic probe-based, real-time PCR. However, the 5'-nuclease activity is substrate dependent and its consequences remain largely unexplored, except for its role in 5'-nuclease-based TaqMan assays. Using both kinetic studies and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), we comprehensively examined the 5'-nuclease activity of Taq DNA polymerase on fluorogenic double-stranded probes of varied structures. We observed that double-stranded probes with destabilized T-terminal could be hydrolyzed, and the major cleavage was the removal of the 5'-terminal fluorophore-labeled nucleotide. These observations can serve as guidance for better design of double-stranded probes with reduced or no interfering background for real-time PCR detection. (C) 2009 Elsevier Ltd. All rights reserved.
语种	英语
出版者	ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD
内容类型	期刊论文
源URL	[http://dx.doi.org/doi:10.1016/j.mcp.2009.04.002]
专题	生命科学－已发表论文
推荐引用方式 GB/T 7714	Huang, Qiuying,Li, Qingge,Li QG. Characterization of the 5 ' to 3 ' nuclease activity of Thermus aquaticus DNA polymerase on fluorogenic double-stranded probes[J],2009.
APA	Huang, Qiuying,Li, Qingge,&李庆阁.(2009).Characterization of the 5 ' to 3 ' nuclease activity of Thermus aquaticus DNA polymerase on fluorogenic double-stranded probes..
MLA	Huang, Qiuying,et al."Characterization of the 5 ' to 3 ' nuclease activity of Thermus aquaticus DNA polymerase on fluorogenic double-stranded probes".(2009).

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