Kinahan David J, Dalton Tara M, Davies Mark R D
Stokes Bio Ltd., Henry St., Limerick, Ireland.
Biomed Microdevices. 2009 Aug;11(4):747-54. doi: 10.1007/s10544-009-9288-6.
In recent years, Fluorescent Melting Curve Analysis (FMCA) has become an almost ubiquitous feature of commercial quantitative PCR (qPCR) thermal cyclers. Here a micro-fluidic device is presented capable of performing FMCA within a microchannel. The device consists of modular thermally conductive blocks which can sandwich a microfluidic substrate. Opposing ends of the blocks are held at differing temperatures and a linear thermal gradient is generated along the microfluidic channel. Fluorescent measurements taken from a sample as it passes along the micro-fluidic channel permits fluorescent melting curves to be generated. In this study we measure DNA melting temperature from two plasmid fragments. The effects of flow velocity and ramp-rate are investigated, and measured melting curves are compared to those acquired from a commercially available PCR thermocycler.
近年来，荧光熔解曲线分析（FMCA）几乎已成为商用定量聚合酶链反应（qPCR）热循环仪的普遍特性。本文展示了一种能够在微通道内进行FMCA的微流控装置。该装置由模块化导热块组成，这些导热块可夹在微流控基板之间。导热块的相对两端保持在不同温度，从而沿微流控通道产生线性热梯度。当样品沿微流控通道通过时进行荧光测量，可生成荧光熔解曲线。在本研究中，我们测量了两个质粒片段的DNA熔解温度。研究了流速和升温速率的影响，并将测得的熔解曲线与从市售PCR热循环仪获得的曲线进行了比较。
