Abstract: The present invention is directed to a method of determining the concentration of a nucleic acid product that had been amplified through polymerase chain reaction (PCR). More particularly, the present invention relates to a method wherein a rate constant is determined for a known concentration of amplified product by monitoring the rate of hybridization of the known concentration, and then the concentration of an unknown concentration of a nucleic acid product can be determined by determining the rate of annealing for the unknown concentration, and calculating the concentration from the rate of annealing and the rate constant.

Abstract: Methods of monitoring hybridization during polymerase chain reaction are disclosed. These methods are achieved with rapid thermal cycling and use of double stranded DNA dyes or specific hybridization probes. A fluorescence resonance energy transfer pair comprises fluorescein and Cy5 or Cy5.5. Methods for quantitating amplified DNA and determining its purity are carried out by analysis of melting and reannealing curves.

Abstract: A thermal cycling device having a sample chamber whose temperature can be rapidly and accurately modulated over a range of temperatures needed to carry out a number of biological procedures, such a the DNA polymerase chain reaction. Biological samples are placed in glass microcapillary tubes and then located inside the sample chamber. A programmable controller regulates the temperature of the sample inside the sample chamber. Once a heating cycle is completed, the controller opens a door to the chamber for venting hot air out and cool ambient air is moved in. Temperature versus time profiles corresponding to optimum denaturation, annealing and elongation temperatures for amplification of DNA are achievable by the present invention.