Methods of determining the presence of double stranded nucleic acids in a sample
AssigneeThe Regents of the University of California
InventorDavid W. Deamer
Life of this patent
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Abstract
Methods for determining the presence of double stranded nucleic acids in a sample are provided. In the subject methods, nucleic acids present in a fluid sample are translocated through a nanopore, e.g. by application of an electric field to the fluid sample. The current amplitude through the nanopore is monitored during the translocation process and changes in the amplitude are related to the passage of single- or double-stranded molecules through the nanopore. The subject methods find use in a variety of applications in which the detection of the presence of double-stranded nucleic acids in a sample is desired, e.g. in hybridization assays, such as Northern blot assays, Southern blot assays, array based hybridization assays, etc.
Claims14
A method of detecting the presence of double stranded nucleic acid molecules in a sample that includes both double and single stranded nucleic acids said method comprising:
- (a) contacting said sample with a single nanopore:
- (b) translocating at least a portion of the nucleic acids present in said sample through said nanopore;
- (c) monitoring the current amplitude through said nanopore during said translocating;
- and
- (d) relating any changes in said current amplitude during said translocating to the presence or absence of double stranded nucleic acid molecules in said sample, wherein said relating comprises distinguishing signals produced by double stranded and single stranded nucleic acids;
- to detect the presence of double stranded nucleic acid molecules in said sample that includes both double and single stranded nucleic acids.
A method of determining the relative amounts of single and double stranded DNA molecules in a sample, said method comprising:
- (a) contacting said aqueous sample with a nanopore device comprising a barrier that includes a single nanopore;
- (b) translocating substantially all DNA molecules present in said sample through said nanopore by applying an electric field to said sample:
- (c) monitoring the current amplitude through said nanopore during said translocating and deriving a current blockade profile;
- and
- (d) relating said current blockade profile to the relative amounts of single and double stranded DNA molecules in said sample;
- to determine the relative amounts of single and double stranded DNA molecules in said sample.
A method of quantitatively determining the amounts of single and double stranded DNA molecules in a sample, said method comprising:
- (a) contacting said aqueous sample with a nanopore device comprising a barrier that comprises a single nanopore;
- (b) translocating substantially all DNA molecules present In said sample through said nanopore by applying an electric field to said sample;
- (c) monitoring the current amplitude through said nanopore during said translocating to obtain a plurality of current amplitude measurements and deriving current blockade profiles from said plurality of current amplitude measurement;
- and
- (d) relating said current blockade profiles to the quantitative amounts of single and double stranded DNA molecules in said sample;
- to detect the quantitative amounts of single and double stranded DNA molecules in said sample
- whereby the quantitative amounts of single and double stranded DNA molecules in said sample are detected.
Dependencies are inferred from the claim text.