I have to perform a 2-step qRT-PCR & I have also constructed a set of in vitro transcribed RNA. My question is, do I make cDNA from the in vitro transcribed RNA & serial dilute the cDNA to plot a standard curve? I am quite lost because the protocols I've read so far either used plasmid DNA or RNA to plot standard curves. I have a problem with my plasmid DNA - the concentration is very low. Help needed please.
Thanks in advance.