3' RACE with just 10ng starting total RNA - (Mar/04/2008 )

Hi everyone,

First time posting on this forum.

I have a tumor sample straight from a clinical patient so it is precious...and my boss will let me take only 10ng (MAYBE 50ng) of total RNA.

I was planning on doing 3' RACE with the SuperScript III kit instead of buying a kit, but I don't know how good it will work with just 10-50ng of total RNA.

Has anyone done an RT reaction with so little starting RNA? Is there a kit on the market that can work at such small concentrations?

It seems like a waste to buy a kit just for 3' RACE, but I will if it is advertised to work with small volumes.

Thanks!

Dilute some non-critical RNA samples to 10 ng and practice multiple times until you are sure things work. Only then move on to your precious sample.

Thanks for the suggestion, phage!

I didn't think of just practicing it...I'll try it today. =)