METHOD FOR PROCESSING POLYNUCLEOTIDE-CONTAINING SAMPLES
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Abstract
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.
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41 Claims
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1-20. -20. (canceled).
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21. A method for processing a polynucleotide-containing sample, the method comprising:
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retaining polynucleotides from a sample on a plurality of binding particles in a process chamber under a first set of conditions, wherein the retaining step comprises binding the polynucleotides to the surfaces of the plurality of binding particles comprising a poly-cationic substance, wherein the sample has a volume from 0.5 microliters to 3 milliliters; wherein the first set of conditions includes a first pH of about 8.5 or less and a first temperature, wherein the first temperature is between about 30°
C. and about 50°
C.;releasing the polynucleotide from the plurality of binding particles under a second set of conditions; and wherein the second set of conditions includes increasing the pH to a second pH by addition of a hydroxide solution and increasing the temperature to a second temperature, wherein the second temperature is between about 80°
C. and about 100°
C. - View Dependent Claims (22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32)
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33. A method for processing a polynucleotide-containing sample, the method comprising:
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retaining polynucleotides from a sample on a plurality of binding particles in a process chamber under a first set of conditions, wherein the retaining step comprises binding the polynucleotides to the surfaces of the plurality of binding particles comprising a poly-cationic substance, wherein the sample has a volume from 0.5 microliters to 3 milliliters; wherein the first set of conditions includes a first pH of about 8.5 or less and a first temperature, wherein the first temperature is about 60°
C.;releasing the polynucleotide from the plurality of binding particles under a second set of conditions; and wherein the second set of conditions includes increasing the pH to a second pH by addition of a hydroxide solution and increasing the temperature to a second temperature, wherein the second temperature is between about 80°
C. and about 100°
C. - View Dependent Claims (34, 35, 36, 37, 38, 39, 40, 41)
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Specification