Cartridge-based thermocycler
Cartridge-based thermocyclers can include cartridges that are configured to move a fluid between distinct chambers. In some cases, the cartridge-based thermocyclers can be used for thermocycling a sample fluid comprising a deoxyribonucleic acid (DNA) target to perform polymerase chain reaction (PCR). Individual chambers can be heated, cooled, and/or compressed to mix fluid within the chamber or to propel fluid in the chamber into another chamber. The cartridges can have a laminate construction. The cartridges can be configured to enable multiplexed thermocycling and/or detection.
1. A method for performing polymerase chain reaction (PCR), comprising:
conveying a sample having a starting volume of at least 50 μL into a first chamber portion of an amplification module, the first chamber portion having a first average temperature;
producing a flow of the sample from the first chamber portion to a second chamber portion of the amplification module, the second chamber portion having a second average temperature, the second average temperature different than the first average temperature such that the sample is thermally cycled when moved from the first chamber portion to the second chamber portion, the amplification module being constructed from a plurality of layers to define the first chamber portion and the second chamber portion, the plurality of layers including at least a top layer, a bottom layer, a first adhesive layer coupled to the top layer, and a second adhesive layer coupled to the bottom layer, wherein at least one layer selected from the group consisting of the top layer and the bottom layer, comprises poly (4,4′-oxydiphenylene-pyromellitimide); and
completing the PCR of the sample by repeatedly flowing the sample between the first chamber portion and the second chamber portion.
2. The method of claim 1 , further comprising completing the PCR amplification within a total thermocycling time of less than about 10 minutes.
3. The method of claim 1 , further comprising filling the first chamber portion with fluid from an ancillary chamber.
4. The method of claim 1 , further comprising detecting PCR amplification by monitoring the first chamber portion, the second chamber portion or a channel between the first chamber portion and the second chamber portion.
5. The method of claim 1 , further comprising heating the first chamber portion or the second chamber portion using one-sided heating.
6. The method of claim 1 , further comprising heating the first chamber portion or the second chamber portion using two-sided heating.
7. The method of claim 1 , wherein the PCR efficiency is at least about 92%.
8. The method of claim 1 , wherein the sample is flowed between the first chamber portion and the second chamber portion at least 20 times.
9. The method of claim 1 , wherein the sample is flowed between the first chamber portion and the second chamber portion at least 30 times.
10. The method of claim 1 , wherein the sample is flowed between the first chamber portion and the second chamber portion at least 40 times.
11. The method of claim 1 , wherein the sample is cycled from the first chamber portion to the second chamber portion and back to the first chamber portion with a cycle time of less than about 15 seconds.
12. The method of claim 11 , wherein the cycle time is less than about 10 seconds.
13. A method for performing polymerase chain reaction (PCR), comprising:
conveying a sample having a starting volume of at least 50 μL into an amplification module having a first portion having and a second portion, the amplification module being constructed from a plurality of layers to define the first portion and the second portion, the plurality of layers including at least a top layer, a bottom layer, a first adhesive layer coupled to the top layer, and a second adhesive layer coupled to the bottom layer, wherein at least one layer selected from the group consisting of the top layer and the bottom layer, comprises poly (4,4′-oxydiphenylene-pyromellitimide);
heating the first portion of the amplification module to a first average temperature;
heating the second portion of the amplification module to a second average temperature, the second average temperature different than the first average temperature;
producing a flow of the sample from the first portion to the second portion of the amplification module such that the sample is thermally cycled when moved from the first portion to the second portion, and such that the sample transitions from the first average temperature to substantially the second average temperature at a rate of 10 μL° C./second or more; and
completing the PCR of the sample by repeatedly flowing the sample between the first portion and the second portion at least 20 times.
14. The method of claim 13 , wherein the sample is flowed between the first portion and the second portion at least 30 times.
15. The method of claim 13 , wherein the sample is flowed between the first portion and the second portion at least 40 times.
16. The method of claim 13 , further comprising completing the PCR amplification within a total thermocycling time of less than about 4 minutes.
17. The method of claim 13 , further comprising filling the first portion with fluid from an ancillary chamber.
18. The method of claim 13 , further comprising detecting PCR amplification by monitoring any of the first portion of the amplification module, the second portion of the amplification module, or a channel between the first portion and the second portion.
19. The method of claim 13 , further comprising heating the first portion of the amplification module or the second portion of the amplification module using one-sided heating.
20. The method of claim 13 , wherein the PCR efficiency is at least about 92%.
21. The method of claim 19 , wherein the amplification module includes a first heater and a second heater, the first heater coupled to a substrate and aligned with the first portion, the second heater coupled to the substrate and aligned with the second portion, the second heater separated from the first heater by an air gap.
22. The method of claim 13 , wherein the first portion and the second portion are fluidically coupled via a channel through which the sample flows from the first portion to the second portion.