Microfluidic system for amplifying and detecting polynucleotides in parallel
US-2015315631-A1 · Nov 5, 2015 · US
US10710069B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-10710069-B2 |
| Application number | US-201414318160-A |
| Country | US |
| Kind code | B2 |
| Filing date | Jun 27, 2014 |
| Priority date | Nov 14, 2006 |
| Publication date | Jul 14, 2020 |
| Grant date | Jul 14, 2020 |
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The present technology provides for a microfluidic substrate configured to carry out PCR on a number of polynucleotide-containing samples in parallel. The substrate can be a single-layer substrate in a microfluidic cartridge. Also provided are a method of making a microfluidic cartridge comprising such a substrate. Still further disclosed are a microfluidic valve suitable for use in isolating a PCR chamber in a microfluidic substrate, and a method of making such a valve.
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What is claimed: 1. A method of making a microfluidic valve in a microfluidic device comprising a reactor and a window disposed relative to the reactor to allow detection of fluorescence from the reactor, the method of making comprising: directing a dispensing head over an inlet hole in a microfluidic substrate of the microfluidic device; propelling a quantity of thermally responsive substance as a succession of fluid dots from the dispensing head into the inlet hole; and maintaining a temperature of the microfluidic substrate so that the thermally responsive substance flows by capillary action from the inlet hole into a microfluidic loading channel in communication with the inlet hole, the microfluidic loading channel comprising a first end and a second end, the second end of the microfluidic loading channel intersecting a microfluidic flow channel at a T-junction such that the flow of thermally responsive substance stops when the thermally responsive substance reaches the microfluidic flow channel without entering the microfluidic flow channel to form the microfluidic valve. 2. The method of claim 1 , wherein the T-junction intersection between the microfluidic loading channel and the microfluidic flow channel comprises a higher cross section than the microfluidic loading channel. 3. The method of claim 1 , wherein capillary forces in the T-junction intersection between the microfluidic loading channel and the microfluidic flow channel are lower than in the microfluidic loading channel. 4. The method of claim 1 , wherein the quantity of thermally responsive substance dispensed by the dispensing head varies between a minimum and a maximum shot size. 5. The method of claim 4 , wherein the volume of the microfluidic loading channel does not exceed the maximum shot size. 6. The method of claim 4 , wherein the quantity is 75±15 nl. 7. The method of claim 1 , wherein the propelling utilizes heated air. 8. The method of claim 1 , wherein the propelling utilizes compressed air. 9. The method of claim 1 , wherein the thermally responsive substance is wax. 10. The method of claim 1 , wherein the dispensing head is heated. 11. The method claim 1 , further comprising maintaining a temperature of the thermally responsive substance in the dispensing head so that the thermally responsive substance is dispensed in molten form into the inlet hole. 12. The method of claim 1 , wherein the directing is automatically controlled. 13. A microfluidic valve made by the method of claim 1 , wherein the microfluidic loading channel and the microfluidic flow channel form a ninety degree angle at the T-junction. 14. A microfluidic substrate comprising one or more microfluidic valves made by the method of claim 1 , wherein the T-junction has a dimension to reduce capillary forces at the T-junction. 15. A method of making a microfluidic valve, the method of comprising: directing a dispensing head over an inlet hole in a microfluidic substrate; propelling a quantity of fluid thermally responsive substance from the dispensing head into the inlet hole; flowing by capillary action the thermally responsive substance from the inlet hole into a microfluidic loading channel in communication with the inlet hole, the microfluidic loading channel comprising a first end and a second end, the second end of the microfluidic loading channel intersecting a microfluidic flow channel; and stopping the flow of thermally responsive substance when the thermally responsive substance reaches the microfluidic flow channel without entering the microfluidic flow channel to form the microfluidic valve. 16. The method of claim 15 , wherein the second end of the microfluidic loading channel intersects the microfluidic flow channel at a T-junction. 17. The method of claim 16 , wherein the T-junction intersection between the microfluidic loading channel and the microfluidic flow channel comprises a higher cross section than the microfluidic loading channel. 18. A method comprising: directing a first dispensing head over a first inlet hole in a microfluidic substrate; propelling a quantity of fluid thermally responsive substance from the first dispensing head into the first inlet hole; flowing by capillary action the thermally responsive substance from the first inlet hole into a microfluidic loading channel in communication with the first inlet hole, the microfluidic loading channel comprising a first end and a second end, the second end of the microfluidic loading channel intersecting a microfluidic flow channel; stopping the flow of thermally responsive substance when the thermally responsive substance reaches the microfluidic flow channel without entering the microfluidic flow channel, wherein the microfluidic substrate comprises a second inlet hole figured to receive a sample dispensed with a second dispensing head. 19. The method of claim 18 , wherein the microfluidic substrate further comprises a plurality of sample lanes configured to receive a plurality of samples, and wherein inlets of the respective plurality of sample lanes are spaced apart from one another to permit simultaneous loading from a multiple-pipette head dispenser.
Disc shape · CPC title
phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers · CPC title
using resistive heater · CPC title
with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples · CPC title
fluid pressure, pneumatics · CPC title
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