I plan to reverse transcribe mRNA from my gene of interest into cDNA then clone it. Starting material will be total RNA from normal cells expressing my gene of interest. I am going to use a high fidelity reverse transcriptase (i.e. recombinant with proofreading activity).
Question: For making cDNA to clone, Is it better to use gene specific primer in this case or use Poly T? I have always used PolyT/random hexamer mix because I wanted total RNA copied to cDNA but not sure in this case. Definitely do not need random hexamers. Any suggestions or comments about this? I am not sure if it really matters because either way I will end up performing PCR on the cDNA to amplify and clone it.
Also, the ORF for this gene is 5kb.