what's wrong with my 3' RACE - (Aug/21/2006 )
I've been stuck with my 3'race for a long time.currently, I use the smart race kit from BD. I did it according to the protocal, but after the 1st round pcr, there was no band---only smear. So I turned to the 2rd nest pcr. This time, it seemed that there were several bands ,but not so clear.
1st round pcr
2rd nest pcr
I'm not sure if I have to start again(extracting RNA------3'RACE), or set a series of pcr reaction using the 1st pcr product as template.
for my 3'RACE i often took 1ul of the first round PCR- diluted it 1:100 and redid the pcr....seemed to get better bands (but my smear wasn;t quite as bright as yours) - how many cycles have you been doing?