THERMAL CYCLING METHOD
First Claim
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1. A method for performing one or more nucleic acid amplification reactions, comprising:
- a. providing one or more sample vessels, each one or more sample vessel comprising a reaction mixture comprising a polynucleotide sample, reagents for carrying out nucleic acid amplification, and a dye for detecting amplification;
b. cycling the temperature of the reaction mixture in the one or more sample vessels to perform multiple amplification cycles, wherein the one or more sample vessels are each in thermal contact with a liquid contained within a heat block wherein the liquid is actively mixed;
c. regulating the temperature of the reaction mixture in each of the one or more sample vessels by heating and cooling the liquid; and
d. optically measuring the dyes between or during each of the multiple amplification cycles to determine a level of amplification while the one or more sample vessels remain in thermal contact with the liquid.
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Abstract
This invention provides a method for carrying out nucleic acid amplification reactions involving heating and cooling of samples in sample vessels utilizing a heat block comprising a liquid. The method can be used to perform multiple nucleic acid amplification reactions simultaneously in which each of the reactions is performed so as to have temperature profiles. The apparatus can be used for performing PCR, and real time PCR in particular, with control and uniformity.
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Citations
24 Claims
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1. A method for performing one or more nucleic acid amplification reactions, comprising:
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a. providing one or more sample vessels, each one or more sample vessel comprising a reaction mixture comprising a polynucleotide sample, reagents for carrying out nucleic acid amplification, and a dye for detecting amplification; b. cycling the temperature of the reaction mixture in the one or more sample vessels to perform multiple amplification cycles, wherein the one or more sample vessels are each in thermal contact with a liquid contained within a heat block wherein the liquid is actively mixed; c. regulating the temperature of the reaction mixture in each of the one or more sample vessels by heating and cooling the liquid; and d. optically measuring the dyes between or during each of the multiple amplification cycles to determine a level of amplification while the one or more sample vessels remain in thermal contact with the liquid. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24)
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Specification