5' RACE Specificity Problems

[Sarah Childs]

I'm having trouble with specificity in 5'RACE.  

I'm getting non-specific bands, even though my gene-specific 
primer is an 18 mer with a Tm of 58C, I'm doing the PCR with a hot start and 
annealing at 56C.  At the same time I am not getting the product I want.  
Has anyone had similar problems and overcome them?  Does anyone have a 
very good RACE protocol that might help me?

Sarah Childs
Ontario Cancer Institute

email: childs at oci.utoronto.ca