When performing a standard curve for some new primers to test for fold change, is it necessary to run the standard curve? I mean, in case your experiment has several genotypes for the same ecotype background, with correspondent control and treatments, must I run the standard curves for all this scenarios, or rather may I pick, just to say a random genotype of treated ones, see that it R=0.99 is or higher and efficiency 2? It is in order to not to waste material.
Do the standard curve once for each primer pair to determine efficiencies. You can use the efficiency values for any number of replicates with any template.
I think we have somewhat discussed this issue in your previous post.