Dynamic flux nucleic acid sequence amplification
First Claim
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1. A real-time dynamic flux method of nucleic acid sequence amplification, comprising:
- a. combining a pair of forward and reverse oligonucleotide primers with a target nucleic acid sequence to be amplified; and
b. amplifying the target nucleic acid sequence by thermocycling the pair of forward and reverse oligonucleotide primers and the target nucleic acid sequence within a 15°
C. temperature range that is defined by the area contained within the overlap of an annealing curve (TA) of the pair of oligonucleotide primers and the denaturation curve (TD) of the target nucleic acid sequence,wherein each forward and reverse oligonucleotide primer has a melting temperature (Tm) within 15°
C. of the Tm of the target nucleic acid sequence, andwherein thermocycling comprises;
i. denaturing the target nucleic acid sequence; and
ii. annealing of the forward and reverse oligonucleotide primers; and
iii. extension of the target nucleic acid sequence by the forward and reverse oligonucleotide primers,c. simultaneously detecting the amplified target nucleic acid sequence during said amplifying step.
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Abstract
Provided herein are dynamic flux nucleic acid sequence amplification methods. The dynamic flux nucleic acid sequence amplification methods described herein are capable of amplifying nucleic acid sequences within a narrow temperature range. In some aspects, the disclosure provides for real-time dynamic flux nucleic acid sequence amplification methods.
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Citations
15 Claims
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1. A real-time dynamic flux method of nucleic acid sequence amplification, comprising:
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a. combining a pair of forward and reverse oligonucleotide primers with a target nucleic acid sequence to be amplified; and b. amplifying the target nucleic acid sequence by thermocycling the pair of forward and reverse oligonucleotide primers and the target nucleic acid sequence within a 15°
C. temperature range that is defined by the area contained within the overlap of an annealing curve (TA) of the pair of oligonucleotide primers and the denaturation curve (TD) of the target nucleic acid sequence,wherein each forward and reverse oligonucleotide primer has a melting temperature (Tm) within 15°
C. of the Tm of the target nucleic acid sequence, andwherein thermocycling comprises; i. denaturing the target nucleic acid sequence; and ii. annealing of the forward and reverse oligonucleotide primers; and iii. extension of the target nucleic acid sequence by the forward and reverse oligonucleotide primers, c. simultaneously detecting the amplified target nucleic acid sequence during said amplifying step. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
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Specification