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q.c. RT-PCR in fishes

Fabio Di Benedetto FabioDB at katamail.com
Thu Feb 13 04:38:07 EST 2003

I'm a simple Biomedical Laboratory Technician, i've some experiences about
PCR for diagnosis of some Human dieses, but now i'm going to start to work
in a laboratory of fish patology and  i know  almost nothing about fishes.
My new duty is to set up a protocoll for diagnosis with RT-PCR of Nervous
Necrosis Virus (VNN) from encephal of Dicentrarcus labrax. "It's not very
difficolt" probably becouse there are many papers about it, but i've a big
problem for the quality control of capacity to amplify (to elongate) of the
RNAs extracteted from the samples .
I need of sequensis of RNA of Dicentrarchus l. that can be used for a
amplify control of all samples of encephal. I hope to do a multiplex with
the primers of the controll and the primers of VNN RNA together but if isn't
possible i can anyway to plan 2 separate PCRs.
It's important to reamark that the  primers of VNN of Dicentrarchus can be
used also for VNN of Umbina cirrosa and Sparus aurata so a control that can
be used also for this fishes would be perfect .
I hope to be clear and that someone that work with Zebra fishes (or with
fishes or with knowledges about RT-PCR) will be able to help me.

Fabio Di Benedetto
FabioDB at katamail.com


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