Pressure-controlled nucleic acid hybridization
AssigneeBBI BioSeq, Inc., Massachusetts corporation
InventorsRobert A. Hess, James A. Laugharn JR., David J. Green
Life of this patent
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Abstract
A method of hybridizing a first nucleic acid to a second nucleic acid at least partially complementary to the first nucleic acid by (1) providing a sample vessel and pressure controller for the vessel; and (2) contacting the first and second nucleic acids within the vessel at a pressure above ambient pressure that is effective to enhance hybridization of the first and second nucleic acids.
Claims16
A method of hybridizing a first nucleic acid to a second nucleic acid at least partially complementary to the first nucleic acid, the method comprising:
- (1) providing a sample vessel and pressure controller for the vessel;
- and
- (2) contacting the first and second nucleic acids within the vessel at a pressure above ambient pressure that is effective to enhance hybridization of the first and second nucleic acids.
A method of detecting in a sample the presence of a nucleic acid that hybridizes to a reference nucleic acid at a first higher pressure but not at a second lower pressure, the method comprising:
- (1) providing a sample vessel and pressure controller for the vessel;
- and in any order
- (2) contacting the reference sequence with the sample in the vessel at the first pressure;
- (3) contacting the reference sequence with the sample in the pressure vessel at the second pressure;
- and
- (4) detecting the presence of a nucleic acid that hybridizes to the reference nucleic acid at the first pressure but not at the second pressure.
A method of discriminating between a first nucleic acid and a second nucleic acid that is different from the first nucleic acid, the method comprising,
- (1) providing a sample vessel and pressure controller for the vessel;
- (2) maintaining the vessel at a constant pressure;
- (3) providing the first and second nucleic acid and a reference nucleic acid in the vessel under conditions that do not allow either the first or the second nucleic acid to hybridize to the reference nucleic acid;
- (4) perturbing at least one condition to establish conditions that permit the first nucleic acid to form a complex with the reference nucleic acid at equilibrium and to permit the second nucleic acid to form a complex with the reference nucleic acid at equilibrium;
- and
- (5) comparing the time necessary to achieve equilibrium hybridization between the first nucleic acid and the reference nucleic acid with the time necessary to achieve equilibrium hybridization between the second nucleic acid and the reference nucleic acid, wherein the difference indicates the relative difference in sequence between the first and the second nucleic acids.
A method of discriminating between a first nucleic acid and a second nucleic acid that is different from the first nucleic acid, the method comprising,
- (1) providing a sample vessel and pressure controller for the vessel;
- (2) providing the first and second nucleic acid and a reference nucleic acid in the vessel under a first pressure that does not allow either the first or the second nucleic acid to hybridize to the reference nucleic acid;
- (3) perturbing the pressure to establish conditions that permit the first nucleic acid to form a complex with the reference nucleic acid at equilibrium and to permit the second nucleic acid to form a complex with the reference nucleic acid at equilibrium;
- and
- (4) comparing the time necessary to achieve equilibrium hybridization between the first nucleic acid and the reference nucleic acid with the time necessary to achieve equilibrium hybridization between the second nucleic acid and the reference nucleic acid, wherein the difference indicates the relative difference in sequence between the first and the second nucleic acids.
Dependencies are inferred from the claim text.