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Real-time PCR |
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| RT-PCR (reverse transcription-polymerase chain reaction) is a sensitive method for the detection of mRNA expression levels. Traditionally RT-PCR involves two steps: the RT reaction and PCR amplification. RNA is first reverse transcribed into cDNA using a reverse transcriptase as described here, the resulting cDNA is used as templates for subsequent PCR amplification using primers specific for one or more genes. RT-PCR can also be carried out as one-step RT-PCR in which all reaction components are mixed in one tube prior to starting the reactions. Although one-step RT-PCR offers simplicity and convenience and minimizes the possibility for contamination, the resulting cDNA can’t be repeatedly used as in the two-step RT-PCR. |
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