VIP in rabbit

[bassanhm]

Emad Gharavi wrote:
> 
> Bassan Haim & Merav:
> 
> Your rat primers to VIP cDNA should work.  In case your RT-PCR needs tuning up
> for this particular signal try using the primers on a rat source of VIP mRNA.
> My physiology isn't so great, so I can't suggest a particular tissue or cell
> line, but my point is that you probably need to make sure you can detect rat
> VIP mRNA from rat tissue (or cells) using your primers before you can start
> with rabbit.  I assume your RNA purification is good and that external
> controls (such as beta actin or GAPDH) primers work fine on your rabbit.
> Can't imagine otherwise, but if not mail me if you need more help.
> 
> Good luck
> 
> E Gharavi
> Emad_G at msn.com

To Emad Gharavi:

Thank you for your answer.

We did get amplification by using the rat primers on rat mRNA.
The major problem with amplification of VIP by mimic RT-PCR is that the
VIP is a 28 amino-acid peptide, in order to succeed in getting mimic
RT-PCR we need to use one primer that is located in the non-translated
area (3`), and it seems  that it might be the problem.
Did you use a particular pair of rat primers that gave an amplification
in rabbit? Or do you have any other idea?

Thank you
Haim & Merav Bassan