URGENT Help !!!

Arturo Sánchez asanchez at cibnor.mx
Wed Jun 4 17:18:20 EST 1997


In recent days I finished some DNA extraction from some marine yeast 
strains, for my thesis work. I am using the electrophoretic patterns as a 
taxonomic tool to identifie marine yeast.
The problem is that I have both DNA genomic and mitochondrial. So, when I 
use some  restriction enzymes (Eco RI, Hinf I, Kpn I, Alu I, and Pst I) I 
can see that in my 2% agarose gel there is some bands, I mean there is 
some patterns, but the backgound is very "strong". Such a high background 
that the paterns are not completly clear. By the way, I must say that in 
a 0.8% gel I have a very clean DNA (only one band, no background).
I have read several articles and always the pictures are very nice, 
almost no background.
 The questions are: 

1) The background is the result of both kinds of DNA? Should I separete 
them by sacarose or CsCl gradients, and then make the restriction?

2) How can I make my gels cleaner? What should I do to have gels good 
enough for pictures?

I am close to finish my thesis, in fact this are my last experiments. So, 
I need an urgent help. I'll be very glad to receive some ideas. Please 
answer to asanchez at cibnor.mx

Thanks a lot.

Arturo Sanchez
Marine Pathology Unit.
Marine Yeast Laboratory.
Center for Biological Research.

La Paz, Baja California Sur. Mexico.




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