METHOD AND DEVICE FOR DETECTING THE PRESENCE OF A SINGLE TARGET NUCLEIC ACID IN A SAMPLE
First Claim
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1. A method for amplifying a target nucleic acid present in a sample, the method comprising:
- introducing into a common fluidic volume a sample and a nongaseous fluid that is substantially immiscible with the sample, wherein the sample comprises reagents sufficient to perform a primer-based amplification of a target nucleic acid when present;
segregating the sample into a plurality of reaction volumes ranging from 1 picoliter to 1 microliter, the reaction volumes being segregated and entrapped by the nongaseous fluid in the common fluidic volume; and
subjecting the plurality of reaction volumes in the common fluidic volume to amplification conditions thereby amplifying the at least one target nucleic acid when present.
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Abstract
A method comprising subjecting one or more sample portion(s) to a single amplification step, thereby amplifying a single molecule in the sample portion to a detectable level, and, in some embodiments, then determining whether the sample portion contains at least one molecule of the target nucleic acid. In some embodiments, the sample portion is in a porous sample structure, or in a sample chamber which comprises means for minimizing diffusion of the sample portion, or in a sample chamber which is inside a microcapillary device, or in a sample retaining means.
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9 Claims
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1. A method for amplifying a target nucleic acid present in a sample, the method comprising:
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introducing into a common fluidic volume a sample and a nongaseous fluid that is substantially immiscible with the sample, wherein the sample comprises reagents sufficient to perform a primer-based amplification of a target nucleic acid when present; segregating the sample into a plurality of reaction volumes ranging from 1 picoliter to 1 microliter, the reaction volumes being segregated and entrapped by the nongaseous fluid in the common fluidic volume; and subjecting the plurality of reaction volumes in the common fluidic volume to amplification conditions thereby amplifying the at least one target nucleic acid when present. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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Specification