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Amplification of a DNA sample via successive rounds of thermal cycling/annealing reactions produces an enormous increase of sample available for measurement. In many instruments for rapid determination, conditions are optimized such that the threshold amplification — and therefore sample detection — occurs after minimal cycling. In different PCR methods the amplified DNA sample can either be the end-point of the determination in a diagnostic application or the amplified sample may be transferred to a sequencing instrument for determination of the actual sequence of DNA bases.
The amplification of a sample of interest and typically no quantification allow for less stringent light detection requirements. Essentially, detection reduces to presence or absence of the signal. However, with ever-increasing demands for faster and greater capacity sequencing instruments, the microtiter plates on which the PCR reactions occur have increased in density and decreased in volume.
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