Method and apparatus for analysis and sorting of polynucleotides based on size
First Claim
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1. A microfabricated device for sorting reporter-labelled polynucleotide molecules by size, comprising a chip having a substrate into which is microfabricated at least one analysis unit, said analysis unit comprising a main channel, having at one end a sample inlet, having along its length a detection region, and having, adjacent and downstream of said detection region, a branch point discrimination region;
- a plurality of branch channels originating at the discrimination region and in communication with the main channel;
means for passing a continuous stream of solution containing said molecules through said detection region, such that on average only one molecule occupies the detection region at any given time;
means for measuring the level of reporter from each molecule within the detection region; and
means for directing said molecule to a selected branch channel based on said level of reporter.
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Abstract
The invention relates to a microfabricated device and methods of using the device for analyzing and sorting polynucleotide molecules by size.
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24 Claims
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1. A microfabricated device for sorting reporter-labelled polynucleotide molecules by size, comprising
a chip having a substrate into which is microfabricated at least one analysis unit, said analysis unit comprising a main channel, having at one end a sample inlet, having along its length a detection region, and having, adjacent and downstream of said detection region, a branch point discrimination region; -
a plurality of branch channels originating at the discrimination region and in communication with the main channel;
means for passing a continuous stream of solution containing said molecules through said detection region, such that on average only one molecule occupies the detection region at any given time;
means for measuring the level of reporter from each molecule within the detection region; and
means for directing said molecule to a selected branch channel based on said level of reporter. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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11. A method of isolating polynucleotides having a selected size, comprising
A) flowing a continuous stream of solution containing reporter-labeled polynucleotides through a channel comprising a detection region having a selected volume, where the concentration of the molecules in the solution is such that most molecules pass through the detection region one by one, B) determining the size of each molecule as it passes through the detection region by measuring the level of said reporter, C) in said continuous stream of solution, diverting (i) molecules having said selected size into a first branch channel, and (ii) molecules not having said selected size into a second branch channel, and D) collecting polynucleotides diverted into said first branch channel.
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24. A method of sizing polynucleotides in solution, comprising
A) flowing a continuous stream of solution containing reporter-labeled polynucleotides through a microfabricated channel comprising a detection region having a selected volume, where the concentration of the molecules in the solution is such that most molecules pass through the detection region one by one, and B) determining the size of each molecule as it passes through the detection region by measuring the level of said reporter.
Specification