Methods for analysis and sorting of polynucleotides
First Claim
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1. A method of isolating polynucleotide molecules based on size, which method comprises:
- (a) flowing a stream of a solution containing reporter-labeled polynucleotide molecules through an analysis unit, wherein the analysis unit is microfabricated onto the substrate of a chip and comprises a main channel having (i) a polynucleotide sample inlet, (ii) a detection region downstream of the sample inlet, and (iii) a branch point discrimination region adjacent to and downstream of the detection region, wherein on average one polynucleotide molecule at a time is placed within the detection region;
(b) determining the size of each polynucleotide molecule as it passes through the detection region by measuring the level of said reporter;
(c) directing polynucleotide molecules in the flow that have a selected size into a branch channel originating at the branch point discrimination region; and
(d) collecting polynucleotides directed into the branch channel.
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Abstract
The invention relates to a microfabricated device and methods of using the device for analyzing and sorting individual polynucleotides, e.g., by size according to an optical signal measured within a detection region of the device. An optical signal such as fluorescence from a reporter molecule associated with the polynucleotide molecules can be used to determine polynucleotide size or to direct selected polynucleotides into one or more selected branch channels of the device.
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Citations
23 Claims
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1. A method of isolating polynucleotide molecules based on size, which method comprises:
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(a) flowing a stream of a solution containing reporter-labeled polynucleotide molecules through an analysis unit, wherein the analysis unit is microfabricated onto the substrate of a chip and comprises a main channel having (i) a polynucleotide sample inlet, (ii) a detection region downstream of the sample inlet, and (iii) a branch point discrimination region adjacent to and downstream of the detection region, wherein on average one polynucleotide molecule at a time is placed within the detection region;
(b) determining the size of each polynucleotide molecule as it passes through the detection region by measuring the level of said reporter;
(c) directing polynucleotide molecules in the flow that have a selected size into a branch channel originating at the branch point discrimination region; and
(d) collecting polynucleotides directed into the branch channel. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23)
a detector that measures the level of the reporter; and
a flow control that directs the flow of polynucleotide molecules.
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23. The method of claim 1, wherein the flow of polynucleotide molecules is directed by at least one pump and at least one valve.
Specification