MICROFLUIDIC APPARATUS, SYSTEMS, AND METHODS FOR PERFORMING MOLECULAR REACTIONS
First Claim
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1. A method to detect a nucleotide, comprising:
- a) restraining movement of a nucleic acid molecule attached to a single particle using a restriction barrier located within a first channel;
b) contacting the nucleic acid molecule with an exonuclease to release the nucleotide, wherein the nucleotide is a terminal nucleotide; and
c) identifying the released nucleotide by associating the released nucleotide with a surface enhanced Raman spectroscopy-active surface, irradiating the released nucleotide with a detection laser beam and measuring Raman emission from the irradiated nucleotide, thereby detecting the nucleotide.
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Abstract
Disclosed herein are methods, apparatuses, and systems for performing nucleic acid sequencing reactions and molecular binding reactions in a microfluidic channel. The methods, apparatuses, and systems can include a restriction barrier to restrict movement of a particle to which a nucleic acid is attached. Furthermore, the methods, apparatuses, and systems can include hydrodynamic focusing of a delivery flow. In addition, the methods, apparatuses, and systems can reduce non-specific interaction with a surface of the microfluidic channel by providing a protective flow between the surface and a delivery flow.
43 Citations
52 Claims
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1. A method to detect a nucleotide, comprising:
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a) restraining movement of a nucleic acid molecule attached to a single particle using a restriction barrier located within a first channel; b) contacting the nucleic acid molecule with an exonuclease to release the nucleotide, wherein the nucleotide is a terminal nucleotide; and c) identifying the released nucleotide by associating the released nucleotide with a surface enhanced Raman spectroscopy-active surface, irradiating the released nucleotide with a detection laser beam and measuring Raman emission from the irradiated nucleotide, thereby detecting the nucleotide. - View Dependent Claims (2, 3, 4, 5, 46, 47, 48, 51, 52)
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6. A method to determine a nucleotide sequence of a nucleic acid molecule, comprising:
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a) restraining movement of a single particle using a restriction barrier located within a first channel, wherein the nucleic acid molecule is attached to the single particle; b) contacting the nucleic acid molecule with an exonuclease to release a terminal nucleotide; and c) identifying a first released nucleotide and a second released nucleotide by irradiating the first released nucleotide and then the second released nucleotide with light from a detection light source, by associating the first released nucleotide and the second released nucleotide with a surface enhanced Raman spectroscopy-active surface, and measuring Raman emission from the irradiated first released nucleotide and then from the second released nucleotide, thereby determining a nucleotide sequence of the nucleic acid. - View Dependent Claims (7, 8, 9, 49, 50)
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10-45. -45. (canceled)
Specification