Amount of RNA (ng) for good ct values. - (Feb/06/2013 )

I was wondering what is a good amount of RNA to use to get good ct values? I have been using 50ng/ul (total 250ng) per reaction and I am getting slightly high ct values. Any advice would be appreciated.

What cDNA kit are you using?

jerryshelly1 on Wed Feb 6 19:57:21 2013 said:

Sorry for the late response.

Hmmm.. I don't really know the kit I am using to make cDNA. All the reagents I use were taken out of the package. I remember I use random hexamers and denature the RNA then add rnase, dntps, a buffer, and ssi (i think that's the name).

Just double check the recommended amount of RNA (ug) that needs to be added with each reaction. If available, use two or three different specs from a couple different labs to confirm your RNA concentration. That can be the biggest contributing factor.