Nucleic acid amplification using microfluidic devices
The present invention provides microfluidic devices and methods using the same in various types of thermal cycling reactions. Certain devices include a rotary microfluidic channel and a plurality of temperature regions at different locations along the rotary microfluidic channel at which temperature is regulated. Solution can be repeatedly passed through the temperature regions such that the solution is exposed to different temperatures. Other microfluidic devices include an array of reaction chambers formed by intersecting vertical and horizontal flow channels, with the ability to regulate temperature at the reaction chambers. The microfluidic devices can be used to conduct a number of different analyzes, including various primer extension reactions and nucleic acid amplification reactions.
1. A microfluidic peristaltic pump comprising a microfluidic flow channel and a first valve, a second valve, and a third valve, wherein the valves occur in a consecutive order along a length of the flow channel and wherein the valves are operably linked such that a single actuation force closes the valves in sequence thereby creating a pumping action through the channel, wherein the sequence is: the first valve is closed before the second valve is dosed, and the second valve is closed before the third valve is dosed.
2. The microfluidic peristaltic pump of claim 1 wherein the single actuation force results in the following sequence of valve states:
i) the first valve is closed, and the second and third valves are open; and then
ii) the first and second valves are dosed, and the third valve is open; and then
iii) the first, second and third valves are closed; and then
iv) the first valve is open, and the second and third valves are closed; and then
v) the first and second valves are open and the third valve is closed; and then
vi) the first, second and third valves are open.
3. The microfluidic peristaltic pump of claim 1 comprising a control channel, wherein:
the flow channel and the control channel are disposed within a substrate,
each valve occurs at a location where the control channel passes over the flow channel, wherein
the valves that occur in a consecutive order along a length of the flow channel are in the same consecutive order along a length of the control channel, and the length of the control channel is longer than said length of the flow channel.
4. The microfluidic peristaltic pump of claim 3 , wherein each valve comprises a deflectable membrane separating the interior of the flow channel from the interior of the control channel.
5. The microfluidic peristaltic pump of claim 4 , wherein the control channel is “S” shaped.
6. A method of pumping fluid through a flow channel using the microfluidic peristaltic pump of claim 3 , the method comprising applying a high pressure signal to one end of the control channel, thereby causing the high pressure signal to propagate to the first valve, then to the second valve, and then to the third valve, thereby actuating the valves in the order in which they occur along the length of the flow channel.
7. The method of claim 6 , further comprising relaxing pressure at the one end of the control channel, causing a low pressure signal to propagate to the first valve, then to the second valve, and then to the third valve, thereby opening the valves in the order in which they occur along the length of the flow channel.
8. The method of claim 7 comprising multiple cycles of applying a high pressure signal to the one end of the control channel and relaxing pressure at the one end of the control channel.
9. A method of pumping fluid using a microfluidic peristaltic pump,
wherein the microfluidic peristaltic pump comprises a microfluidic flow channel, a first valve, a second valve, and a third valve, the valves being operably linked,
and the method comprises applying a single actuation force to the valves, thereby closing the valves in a dosing sequence, such that the first valve is closed before the second valve is closed, and the second valve is closed before the third valve is closed,
thereby creating a pumping action through the flow channel.
10. The method of claim 9 , wherein
the microfluidic peristaltic pump further comprises a control channel,
each valve occurs at a location where the control channel passes over the flow channel, and
applying a single actuation force to the control channel comprises applying a high pressure signal to one end of the control channel.
11. The method of claim 10 , further comprising removing the single actuation force, thereby opening the valves in an opening sequence, such that the first valve is opened before the second valve is opened, and the second valve is opened before the third valve is opened.