Thermal gradient for fluorometric optimization of droplet PCR in virtual reaction chambers

	Thermal gradient for fluorometric optimization of droplet PCR in virtual reaction chambers
	Li, X. P.; W. M. Wu and A. Manz
	2017
发表期刊	Microchimica Acta
卷号	184 期号:9
摘要	An open system with a thermal gradient is described for the optimization of polymerase chain reactions (PCR). Two thermal electric coolers were used as the heat source. The gradient is measured through encapsulated water-based beads of a temperature-dependent dye inside mineral oil, thereby forming virtual reaction chambers. Nine droplets (with typical volume of 0.7 mu L) were used. Using the intrinsic fluorescence of a temperature-sensitive inert dye (sulforhodamine B), the process involves measurement of the fluorescence intensity at a known, uniform temperature together with the instrument-specific calibration constant to calculate an unknown, possibly non-uniform temperature. The results show that a nearly linear thermal gradient is obtained. This gradient function is a useful feature that can be used for optimization of a commonly used enzyme-activated reaction, viz. PCR. The emission spectra of fluorescent droplets during two-step PCR were monitored and the changes in fluorescence between 50 degrees C and 100 degrees C quantified. As the gradient feature allows for testing a range of annealing temperatures simultaneously, the optimal annealing temperature can be easily determined in a single experiment.
收录类别	sci ; ei
语种	英语
文献类型	期刊论文
条目标识符	http://ir.ciomp.ac.cn/handle/181722/59026
专题	中科院长春光机所知识产出
推荐引用方式 GB/T 7714	Li, X. P.,W. M. Wu and A. Manz. Thermal gradient for fluorometric optimization of droplet PCR in virtual reaction chambers[J]. Microchimica Acta,2017,184(9).
APA	Li, X. P.,&W. M. Wu and A. Manz.(2017).Thermal gradient for fluorometric optimization of droplet PCR in virtual reaction chambers.Microchimica Acta,184(9).
MLA	Li, X. P.,et al."Thermal gradient for fluorometric optimization of droplet PCR in virtual reaction chambers".Microchimica Acta 184.9(2017).

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