Question

how do you make dna using reverse transcriptase?
I am given a strand of RNA and need to find out how to make the DNA (using reverse transcriptase)

Answer 1

Reverse transcription:
RNA ------> cDNA
It is mediated by reverse transcriptase
Reverse transcriptase is an RNA dependnet DNA polymerase.

For reverse transcription reaction (RT-PCR), the following components are required.
1. RNA sample
2. Primer (It can be Oligo dT or gene specific primer)
3. Buffer
4. dNTPS
5. Reverse transcriptase
6. RNase inhibitor

Prepare a reaction mix with the above compoennets and incubate it at the optimum temperature to obtain cDNA.