Methods for nucleic acid isolation and kits using a microfluidic device and sedimenting reagent
First Claim
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1. A method of isolating nucleic acid from a sample, the method comprising:
- providing a microfluidic device comprising a loading chamber, a valved process chamber, and a mixing chamber;
providing a sample comprising nucleic acid-containing material and inhibitors;
providing a sedimenting reagent;
placing the sample in the loading chamber;
transferring the sample to the valved process chamber;
forming a concentrated region of the sample in the valved process chamber using the sedimenting reagent, wherein the concentrated region of the sample comprises a majority of the nucleic acid-containing material and a less concentrated region of the sample comprises at least a portion of the sedimenting reagent and at least a portion of the inhibitors;
activating a valve in the valved process chamber to transfer at least a portion of the concentrated region of the sample to the mixing chamber and separate at least a portion of the concentrated region from the less concentrated region of the sample;
lysing the nucleic acid-containing material with optional heating in the mixing chamber to release nucleic acid; and
optionally adjusting the pH of the sample comprising released nucleic acid.
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Abstract
The present invention provides methods and kits for isolating nucleic acid from a sample, preferably from a biological sample, using a microfluidic device and sedimenting reagent.
95 Citations
13 Claims
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1. A method of isolating nucleic acid from a sample, the method comprising:
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providing a microfluidic device comprising a loading chamber, a valved process chamber, and a mixing chamber;
providing a sample comprising nucleic acid-containing material and inhibitors;
providing a sedimenting reagent;
placing the sample in the loading chamber;
transferring the sample to the valved process chamber;
forming a concentrated region of the sample in the valved process chamber using the sedimenting reagent, wherein the concentrated region of the sample comprises a majority of the nucleic acid-containing material and a less concentrated region of the sample comprises at least a portion of the sedimenting reagent and at least a portion of the inhibitors;
activating a valve in the valved process chamber to transfer at least a portion of the concentrated region of the sample to the mixing chamber and separate at least a portion of the concentrated region from the less concentrated region of the sample;
lysing the nucleic acid-containing material with optional heating in the mixing chamber to release nucleic acid; and
optionally adjusting the pH of the sample comprising released nucleic acid. - View Dependent Claims (12)
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2. A method of isolating nucleic acid from a sample, the method comprising:
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providing a microfluidic device comprising a loading chamber, a valved process chamber, and a mixing chamber;
providing a sample comprising nucleic acid-containing material and cells containing inhibitors;
providing a sedimenting reagent;
placing the sample in the loading chamber;
transferring the sample to the valved process chamber;
forming a concentrated region of the sample in the valved process chamber using the sedimenting reagent, wherein the concentrated region of the sample comprises a majority of the nucleic acid-containing material and a less concentrated region of the sample comprises at least a portion of the sedimenting reagent and at least a portion of the inhibitors;
activating a valve in the valved process chamber to transfer at least a portion of the concentrated region of the sample to the mixing chamber and separate at least a portion of the concentrated region from the less concentrated region of the sample;
lysing the nucleic acid-containing material in the mixing chamber to release nucleic acid; and
optionally adjusting the pH of the sample comprising released nucleic acid. - View Dependent Claims (3, 4, 5, 6, 7, 8, 9, 10, 11, 13)
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Specification