RT-PCR: Two-Step Protocol - MIT OpenCourseWare
We recommend the two-step protocol for this class. In the one-step protocol, the components of RT and PCR are mixed in a single tube at the same time. The one-step protocol generally works well for amplifying targets that are reasonably abundant. Alternatively, RT-PCR can be done in two steps, first with the reverse transcription and then the PCR.
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