The success of a qPCR reaction is evaluated using a standard dilution series, the data from which is used to calculate the reaction efficiency and R2 values. Efficiency can be defined as the increase in amplicon per cycle, whilst R2 is the coefficient of determination. Ideally, the efficiency of a qPCR reaction would be 100%, meaning that the PCR product is exactly doubling every cycle during the logarithmic phase. Criteria are put in place to determine how successful a qPCR protocol is, with an efficiency of 90-110% and an R2 > 0.985 being deemed acceptable. The xxpress® qPCR thermal cycler performs successful, repeatable qPCRs even with low volume reactions, enabling optimal use of reagents.
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