Real-time PCR in micro-channels
The present invention relates to methods for amplifying nucleic acids in micro-channels. More specifically, the present invention relates to methods for performing a real-time polymerase chain reaction (PCR) in a continuous-flow microfluidic system and to methods for monitoring real-time PCR in such systems.
1. A system for performing real-time PCR in a microfluidic device, said system comprising:
a microfluidic device comprising at least one channel configured to contain at least one test solution containing real-time PCR reagents;
a thermal transfer element integral with or proximal to the microfluidic device, said thermal transfer element configured to thermocycle the test solution along a defined section of the at least one channel for performing real-time PCR in the test solution;
a source of illumination integral with or proximal to the microfluidic device and configured to illuminate the at least one channel; and
a detector integral with or proximal to the microfluidic device, said detector configured to simultaneously detect a fluorescent signal at two or more of a plurality of locations along said defined section of said at least one channel, wherein each of the simultaneously measured fluorescent signals at each location along the channel corresponds to the same point in time within a PCR cycle.
2. The system of claim 1 , wherein the fluorescent signal is measured at least once during each PCR cycle.
3. The system of claim 1 , wherein the detector is configured to detect a signal from amplification products and a signal from a marker at one or more of a plurality of locations along said defined section of said at least one channel.
4. The system of claim 3 , wherein the detector is configured to independently detect signals from the amplified product and the marker.
5. The system of claim 1 , wherein the at least one channel comprises a plurality of channels.
6. The system of claim 1 , wherein the at least one test solution comprises a plurality of test solutions.
7. The system of claim 1 , further comprising a sensor to detect the temperature of the thermal transfer element and to provide feedback information for controlling the temperature cycles.
8. The system of claim 3 , further comprising means for analyzing the amplified product.