Accurate Isothermal Amplification Reaction Rate Determination Using HighFrequency Sampling

Nucleic acid amplification can provide sensitive and specific molecular detection and quantification. Since its introduction PCR has been the dominant amplification technology; its widespread use has stimulated improvements in enzymes, instrumentation, and analytical methods. Initial use of PCR was largely qualitative: positive target detection, or inference of target absence above a detection limit. PCR was made more quantitative with the advent of real­time PCR, notably for relative quantification that estimates a target concentration ratio between two samples. More recently, digital PCR (dPCR) has allowed absolute quantification – the estimation of the number of target molecules in a given sample volume.