Microchannel Fluorescent Melting Curve Analysis

In recent years, Fluorescent Melting Curve Analysis (FMCA) has become an almost ubiquitous feature of commercial quantitative PCR (qPCR) thermal cyclers. Here a microfluidic device is presented capable of performing droplet based Fluorescent Melting Curve Analysis 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 be generated. In this study we demonstrate the capability of measuring fluorescent melt curves from DNA passing through the device in a serial flow of sample plugs buffered by mineral oil.