VIP in rabbit
bassanhm at netvision.net.il
Tue May 27 23:56:35 EST 1997
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?
Haim & Merav Bassan
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