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Real-time polymerase chain reaction monitors fluorescence during cycling. With validated specificity, calibration and PCR efficiency, that record supports estimating initial template amount; this quantitative use is called qPCR. Ordinary endpoint PCR amplification examines accumulated product afterwards. qPCR need not start from RNA, and RT-PCR need not monitor fluorescence. Combining both operations gives RT-qPCR.^[ref]

RT-qPCR applies qPCR after reverse transcription. Differences in RNA recovery and conversion into Complementary DNA can affect its inferred starting amount as well as differences in PCR efficiency.