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 . (canceled)
2 . A system comprising:
a tubular vessel comprising an opened first end and a second end, opposite the opened first end;
an actuatable member configured to open, wherein the actuatable member is positioned closer to the second end than the opened first end;
a closure configured to close the opened first end;
a pressure member configured to be disposed within the tubular vessel; and
a region disposed within the tubular vessel and positioned between the opened first end and the second end,
wherein the tubular vessel is configured to receive a polynucleotide-containing sample through the opened first end and into the region, wherein the polynucleotide-containing sample is configured to be heated within the region once the opened first end is closed with the closure, and
wherein the pressure member is configured to expel a solution with released polynucleotides through the second end.
3 . The system of claim 2 , wherein the tubular vessel comprises a pre-loaded reagent within the tubular vessel.
4 . The system of claim 2 , wherein the actuatable member is configured to open and close.
5 . The system of claim 2 , wherein the pressure member is positioned closer to the opened first end than the second end.
6 . The system of claim 2 , wherein the closure is configured to prevent evaporation of the polynucleotide-containing sample.
7 . The system of claim 2 , further comprising an external heat source, wherein the external heat source is configured to apply heat to the tubular vessel.
8 . The system of claim 2 , further comprising an operator disposed about the tubular vessel, wherein the operator is configured to process the polynucleotide-containing sample within the tubular vessel.
9 . The system of claim 2 , further comprising an operator comprising a first heat source, wherein the first heat source is configured to apply heat to the region.
10 . The system of claim 9 , wherein the operator further comprises a second heat source.
11 . The system of claim 9 , wherein the operator further comprises a second heat source configured to apply heat at least partially below the actuatable member.
12 . The system of claim 10 , wherein the first heat source and the second heat source are separated.
13 . The system of claim 10 , wherein the first heat source and the second heat source are configured to apply heat to regions having different lengths.
14 . The system of claim 10 , wherein the first heat source is configured to apply heat to a larger region than the second heat source.
15 . The system of claim 2 , further comprising an intermediate region disposed within the tubular vessel and positioned between the region and the second end.
16 . The system of claim 15 , wherein the intermediate region has a smaller cross-sectional dimension than the region.
17 . The system of claim 2 , wherein the pressure member is configured to expel a predetermined volume.
18 . The system of claim 17 , wherein the predetermined volume is less than 500 microliters.
19 . The system of claim 17 , wherein the predetermined volume is less than 50 microliters.
20 . The system of claim 2 , wherein the closure has a larger cross-sectional dimension than the tubular vessel.
21 . The system of claim 2 , wherein the tubular vessel is configured to receive a polynucleotide-containing sample having a volume from 100 μL to about 2 mL.