Is it possible to synthesize cDNA manually? if yes then what protocol should follow?

05 November 2022 1 159 Report

I don't have a cDNA kit right now. Further, if there is any protocol to synthesize cDNA from mRNA then which equipments will use. I have Dry bath and refrigerated ultracentrifuge.

Govindkumar Balagannavar

Yes, but you need to have all the required chemicals along with your template.

1. M-MuLV Reverse Transcriptase (NEB: M0253S)

2. oligo(dT) or random hexamer mixture or specific primers

3. dNTPs

you can maintain the required incubation at 42 degrees for a 30mins to an hour and enzyme inactivation at 70 degrees for 5-10min using the dry bath.

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