RACE

[Peter Ellis]

When doing a RACE reaction, you need to use high specificity primers with 
a higher annealing temperature than usual, since you're only using one 
gene-specific primer in the PCR (along with the adaptor primer).  The RACE 
kit, adaptor primers etc. are thus optimised for this.

If you're doing two rounds of amplification, then the second round of PCR 
can use primers with a lower Tm, as the first round should have already 
increased the representation of your gene of interest.

Peter Ellis


On Tue, 23 Sep 2008, Scott Brown wrote:

>
> Dpes anyone know why the annealing temperatures for 3'RACE are always
> much higher than normal?
>
> S
>
>
> Children's Cancer Institute Australia for Medical Research is the only independent medical research institute in Australia devoted to research into the causes, prevention, better treatment and ultimately a cure of childhood cancer. Our vision is to save the lives of all children with cancer and to eliminate their suffering.
>
> The information contained in this message and any annexure is confidential and intended only for the named recipient(s). If you have received this message in error please contact the sender immediately and destroy the original message.
>
>