Microfluidic siphoning array for nucleic acid quantification
In some aspects, the present disclose provides methods for amplifying and quantifying nucleic acids. Methods for amplifying and quantifying nucleic acids comprise isolating a sample comprising nucleic acid molecules into a plurality of microchambers, performing a polymerase chain reaction on the plurality of microchambers, and analyzing the results of the polymerase chain reaction. In some aspects, the present disclosure provides devices consistent with the methods herein.
1. A device for processing a biological sample, comprising:
a plurality of chambers configured to retain said biological sample during processing; and
at least one gas permeable film or membrane covering said plurality of chambers, wherein said at least one gas permeable film or membrane is configured to permit gas flow from a chamber of said plurality of chambers to a location external to said chamber.
2. The device of claim 1 , wherein said at least one gas permeable film or membrane is a plurality of gas permeable films or membranes, and wherein said chamber is covered by a gas permeable film or membrane of said plurality of gas permeable films or membranes.
3. The device of claim 1 , wherein said at least one gas permeable film or membrane seals said plurality of chambers.
4. The device of claim 1 , further comprising a fluid flow path in fluid communication with said plurality of chambers, wherein said fluid flow path is configured to provide reagents to said plurality of chambers.
5. The device of claim 4 , further comprising a plurality of siphon apertures in fluid communication with said fluid flow path and said plurality of chambers, wherein said plurality of siphon apertures is configured to provide reagents from said fluid flow path to said plurality of chambers.
6. The device of claim 1 , wherein said at least one gas permeable film or membrane comprises a polymer.
7. The device of claim 1 , wherein said at least one gas permeable film or membrane has a thickness of at most about 100 micrometers.
8. The device of claim 1 , wherein said at least one gas permeable film or membrane has a thickness of less than or equal to about 50 micrometers.
9. The device of claim 1 , wherein said at least one gas permeable film or membrane is configured to permit gas flow therethrough upon application of a pressure differential in excess of a threshold.
10. The device of claim 1 , wherein said at least one gas permeable film or membrane is configured to be impermeable to water.
11. A device for processing a biological sample, comprising a plurality of chambers configured to retain said biological sample during processing, wherein a chamber of said plurality of chambers comprises a barrier that is configured to permit gas flow from a chamber of said plurality of chambers to a location external to said chamber.
12. The device of claim 11 , wherein said barrier is a gas permeable film or membrane.
13. The device of claim 11 , further comprising a fluid flow path in fluid communication with said plurality of chambers, wherein said fluid flow channel is configured to provide reagents to said plurality of chambers.
14. The device of claim 11 , wherein said barrier has a thickness of at most about 100 micrometers.
15. The device of claim 11 , wherein said barrier has a thickness of less than or equal to about 50 micrometers.
16. The device of claim 11 , wherein said chamber comprises a plurality of barriers, and wherein said barrier is separable from one or more other barriers of said plurality of barriers.
17. The device of claim 11 , wherein said barrier comprises a polymer.
18. The device of claim 11 , wherein said barrier is configured to be impermeable to water.
19. The device of claim 11 , wherein said barrier is configured to permit gas flow therethrough upon application of a pressure differential in excess of a threshold.
20. The device of claim 11 , further comprising a fluid flow path in fluid communication with said plurality of chambers, and wherein said fluid flow path is configured to provide reagents to said plurality of chambers.
21. The device of claim 1 , wherein said chamber has a depth of less than or equal to about 500 micrometers.
22. The device of claim 11 , wherein said chamber has a depth of less than or equal to about 500 micrometers.
23. The device of claim 1 , further comprising a pressure unit configured to supply a pressure drop across said at least one gas permeable film or membrane, which pressure drop is sufficient to induce said gas flow from said chamber to said location external to said chamber.
24. The device of claim 11 , further comprising a pressure unit configured to supply a pressure drop across said barrier, which pressure drop is sufficient to induce said gas flow from said chamber to said location external to said chamber.
25. The device of claim 6 , wherein said polymer is an inelastic material.
26. The device of claim 6 , wherein said polymer is a thermoplastic.
27. The device of claim 1 , wherein said at least one gas permeable film or membrane is at least one gas permeable film.
28. The device of claim 1 , wherein said at least one gas permeable film or membrane is at least one gas permeable membrane.
29. The device of claim 17 , wherein said polymer is an inelastic material.
30. The device of claim 17 , wherein said polymer is a thermoplastic.