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  • Steps are: 1) RNA is first incubated with a primer at 70 degree to denature RNA secondary structure and then quickly chill on ice to let the primer anneal to the RNA. 2) Other components of RT are added to the reaction including dNTPs, RNase inhibitor, reverse transcriptase and RT buffer. 3) RT reaction is extended at 42 degree for 1 hr. 4) Heat the reaction at 70 degree to inactivate the enzyme.
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