Methods for quantitating DNA using digital multiple displacement amplification
First Claim
1. A method of quantifying a nucleic acid molecule comprising(a) contacting a sample to be tested with whole-genome amplification (WGA) reaction components and a label that detects nucleic acid molecules to form a reaction sample;
- (b) partitioning the reaction sample at limiting dilution into monodisperse droplets, wherein each droplet corresponds to a single WGA reaction comprising a single nucleic acid molecule or no nucleic acid molecule;
(c) allowing the WGA reaction to occur in the droplets; and
(d) determining the number of droplets comprising nucleic acid molecules by detecting the label, thereby quantifying the nucleic acid in the sample to be tested.
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Accused Products
Abstract
The present invention relates to methods of quantifying nucleic acids involving (a) contacting a sample to be tested with whole-genome amplification (WGA) reaction components and a dye molecule to form a reaction sample, wherein the dye molecule detects the nucleic acid; (b) partitioning the reaction sample wherein each partitioned reaction sample corresponds to a single reaction; (c) allowing the reaction to occur in the partitioned reaction sample and (d) determining the number of partitioned reaction samples having the dye molecule, wherein the dye molecule indicates the presence of the nucleic acid; thereby quantifying the nucleic acid in the sample to be tested.
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Citations
20 Claims
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1. A method of quantifying a nucleic acid molecule comprising
(a) contacting a sample to be tested with whole-genome amplification (WGA) reaction components and a label that detects nucleic acid molecules to form a reaction sample; -
(b) partitioning the reaction sample at limiting dilution into monodisperse droplets, wherein each droplet corresponds to a single WGA reaction comprising a single nucleic acid molecule or no nucleic acid molecule; (c) allowing the WGA reaction to occur in the droplets; and (d) determining the number of droplets comprising nucleic acid molecules by detecting the label, thereby quantifying the nucleic acid in the sample to be tested. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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15. A method of amplifying a nucleic acid molecule in a sample comprising
(a) contacting a sample with whole-genome amplification (WGA) reaction components and a label that detects nucleic acid molecules to form a reaction sample; -
(b) partitioning the reaction sample at limiting dilution into a hydrogel; and (c) allowing the WGA reaction to occur in the hydrogel. - View Dependent Claims (16, 17, 18, 19)
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20. A method of preparing nucleic acid molecules for analysis, the method comprising:
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(a) contacting a sample to be tested with whole-genome amplification (WGA) reaction components and a label that detects nucleic acid molecules to form a reaction sample; (b) partitioning the reaction sample into monodisperse droplets, wherein each droplet corresponds to a single WGA reaction comprising a single nucleic acid molecule or no nucleic acid molecule; and (c) allowing the WGA reaction to occur in the droplets.
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Specification