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.
Marine Pathology Unit.
Marine Yeast Laboratory.
Center for Biological Research.
La Paz, Baja California Sur. Mexico.