RT-PCR problem!!

Olivier Gandrillon Olivier.Gandrillon at cri.ens-lyon.fr
Mon Mar 22 03:09:07 EST 1999


You might actually have too much template to start with. You can try 
either dilute your fisrt RT-PCR product (10 up to one million times) or 
try lower PCR cycles numbers;

Hope this helps.

Olivier Gandrillon

Dr O. Gandrillon
Département de biologie
Ecole Normale supérieure de Lyon
46, allée d'Italie
69364 Lyon Cedex 7
Phone: 04-72-72-86-15
Fax: 04-72-72-80-80
E.mail: ogandril at ens-lyon.fr
W3: http://www.ens-lyon.fr:80/~ogandril
"The reasonable man adapts himself to the world; the unreasonable one 
persists in trying to adapt the world to himself. Therefore, all 
progress depends on the unreasonable man."
George Bernard Shaw

tahar aboulkassim wrote:
> Hi everyone!
> I did an RT-PCR and I had a band with the right size. I purified this band
> from the gel and dissolve it in tris buffer. then I used 1µl of this RT-PCR
> product and run another PCR (All the parameters were the same indeed the
> primers).But I always obtain a smear.
> I have no explanation for that and I will be so grateful if someone gives me
> an idea.
> thank you so much for help.
> A. Taha
> E- mail : ellape at sprint.ca


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