Algae project, need advice please.
acaudy at FREENET.COLUMBUS.OH.US
Tue Nov 29 18:26:18 EST 1994
On Tue, 29 Nov 1994, Richard Dana wrote:
> I am working on purification of DNA and RNA from single celled algae.
> I would appreciate any help that I can get.
> I did a phenol/chloroform/detergent method today but woulf like to know
> what others have done.
> I work for a company in San Diego.
> Richard Dana
> 1070 Joshua Way
> Vista, CA 92083
> e-mail ggreen at electriciti.com
I've never worked with algae, but I was trying a tricky gram+
bacterium for DNA analysis. I tried potassium acetate/detergent and
couldn't find any DNA. It was in the precipitate, of course. It was
messy and yucky, but I dissolved the pellet in H20 and overlaid with
isopropanol. Then, I removed the DNA at the interface, allowed to
reprecipitate, removed DNA (and other junk of course), reprecipitate, etc.
Then I did RNAse, phenyl-chloroform, etc. It seemed to work well,
although the DNA removal at the interface was a pain. (Brief stints in a
-80 degrees C helped speed precipitation.)
Have you tried proteinase K? I don't know whether it would work
on algae. The protocol is in the Gibco BRL manual for PCR, I think.
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