Thermal gradient for fluorometric optimization of droplet PCR in virtual reaction chambers
文献类型：期刊论文
| 作者 | Li, X. P.; W. M. Wu and A. Manz |
| 刊名 | Microchimica Acta
 |
| 出版日期 | 2017 |
| 卷号 | 184期号:9 |
| 英文摘要 | 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. |
| 语种 | 英语 |
| 源URL | [http://ir.ciomp.ac.cn/handle/181722/59026]  |
| 专题 | 长春光学精密机械与物理研究所_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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