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US 9,745,623 B2Granted
Square wave thermal cycling
AssigneeBecton, Dickinson and Company
InventorAdam Steel
Life of this patent
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Abstract
Embodiments disclosed herein relate to methods and systems for analysis of melting temperatures, and particularly to analysis of duplex nucleic acids.
Claims25
A method of determining the melting temperature of a duplex nucleic acid molecule, said method comprising
- subjecting a solution comprising a double-stranded nucleic acid molecule to a square wave temperature gradient, said gradient comprising a series of temperature pulses, and each pulse comprising:
- a) increasing the temperature of said solution to a first high temperature;
- b) decreasing the temperature of said solution to a second low temperature;
- wherein said first high temperature is greater than said second low temperature, and in subsequent pulses, said first high temperature and said second low temperature are each adjusted to be greater than the first and second temperatures, respectively, in prior pulses;
- and wherein at least one of said temperature pulses comprises a range of temperatures that causes at least one transition of said double-stranded nucleic acid molecule to single-stranded nucleic acid molecules;
- and
- detecting said at least one transition over the course of said square wave temperature gradient, wherein said at least one transition comprises the melting temperature of said duplex nucleic acid molecule.
4 more elements
A method of determining melting temperature of a double-stranded nucleic acid molecule, said method comprising:
- providing a sample comprising double-stranded nucleic acid molecules and a reporter moiety having a signature detectable signal in the presence of duplex nucleic acid;
- subjecting said sample to a series of temperature pulses, wherein at least a plurality of said pulses cause said double-stranded nucleic acid molecules to transition to single-stranded molecules and back to double-stranded molecules, wherein each of said temperature pulses comprise:
- a) increasing the temperature of said solution to a first high temperature,
- b) decreasing the temperature of said solution to a second low temperature,
- wherein said first high temperature is greater than said second low temperature, and in subsequent pulses, said first high temperature and said second low temperature are each adjusted to be greater than the first and second temperatures, respectively, in prior pulses;
- and
- detecting, using the signal produced by said reporter moiety, the melting temperature of said double-stranded nucleic acid molecule.
4 more elements
Dependencies are inferred from the claim text.