Method for processing polynucleotide-containing samples
Methods and systems for processing polynucleotides (e.g., DNA) are disclosed. A processing region includes one or more surfaces (e.g., particle surfaces) modified with ligands that retain polynucleotides under a first set of conditions (e.g., temperature and pH) and release the polynucleotides under a second set of conditions (e.g., higher temperature and/or more basic pH). The processing region can be used to, for example, concentrate polynucleotides of a sample and/or separate inhibitors of amplification reactions from the polynucleotides. Microfluidic devices with a processing region are disclosed.
1. A system comprising:
a tubular vessel comprising an opened end and an opening, the opening opposite the opened end of the tubular vessel;
a valve for opening and closing the opening;
a closure configured to close the opened end;
a gas pressure generator disposed within the tubular vessel; and
a region disposed within the tubular vessel and positioned between the gas pressure generator and the opening,
wherein the tubular vessel is configured to receive a polynucleotide-containing sample through the opened end and into the region, wherein the polynucleotide-containing sample is configured to be heated within the region once the opened end is closed with the closure, and
wherein the gas pressure generator is configured to expel a solution with released polynucleotides from the tubular vessel through the opening.
2. The system of claim 1 , wherein the valve is a thermally actuated valve for opening and closing the opening.
3. The system of claim 1 , wherein the tubular vessel comprises a pre-loaded reagent positioned within the tubular vessel.
4. The system of claim 1 , wherein the closure is configured to prevent evaporation of the polynucleotide-containing sample.
5. The system of claim 1 , further comprising an external heat source, wherein the tubular vessel is configured to be heated by the external heat source.
6. The system of claim 1 , further comprising an operator comprising a first heat source, wherein the first heat source is configured to apply heat to the region disposed within the tubular vessel.
7. The system of claim 6 , wherein the operator further comprises a second heat source, wherein the second heat source is configured to apply heat to another region disposed within the tubular vessel.
8. The system of claim 7 , wherein the first heat source and the second heat source are spaced apart.
9. The system of claim 7 , wherein the first heat source and the second heat source have a different length.
10. The system of claim 7 , wherein the first heat source is configured to apply heat to a larger region than the second heat source.
11. The system of claim 1 , wherein the tubular vessel comprises a pre-loaded enzyme positioned within the tubular vessel.
12. The system of claim 1 , wherein the gas pressure generator comprises a heat source to heat gas present within the gas pressure generator.
13. The system of claim 12 , wherein the heated gas is configured to expand and provide the gas pressure to expel the solution.
14. The system of claim 1 , wherein the gas pressure generator is configured to expel a predetermined volume.
15. The system of claim 14 , wherein the predetermined volume is less than 500 microliters.
16. The system of claim 14 , wherein the predetermined volume is less than 50 microliters.
17. The system of claim 1 , wherein the region is tapered.
18. The system of claim 1 , wherein the tubular vessel is tapered toward the opening.
19. The system of claim 1 , wherein the closure has a larger diameter than the tubular vessel.
20. The system of claim 1 , wherein the tubular vessel comprises a wash solution valve configured to release an amount of wash solution within the tubular vessel.