[Taxacom] Recalcitrant DNAs

Dan Lahr dlahr at ib.usp.br
Tue Oct 20 07:28:44 CDT 2015


Hi Eric,

Amoebae are awful for basically all molecular procedures.  It has to be
something about their cellular chemistry that has made our lives difficult
for many years.  I've tried many many things, which I am happy to share -
the most effective protocols have been guanidine thiocyanate for small
numbers of cells, and phenol:chloroform for large cultures.

Eventually, we moved on to RNA and that is pure bliss for amoebae.
Everything simply works.  My suspicion is that this game is a lot different
for plants.

Let me know if you need more details.

Best,

dan

__________________________________
Daniel J. G. Lahr
PhD, Assist. Prof.
Dept of Zoology, Univ. of Sao Paulo, Brazil
Office number: + 55 (11) 3091 0948
http://www.ib.usp.br/zoologia/lahr/


On Mon, Oct 19, 2015 at 11:44 PM, Tepe, Eric (tepeej) <tepeej at ucmail.uc.edu>
wrote:

> I have a student working in my lab who wants to compare DNA extraction
> protocols on plants that are notoriously difficult to sequence.  I am
> looking for any plant species or groups that are difficult to extract for
> whatever reason, have PCR inhibiting chemistry, or are otherwise
> problematic.  The trickier the better!  If you have experience with any of
> these, please let me know.
>
> Thanks in advance,
> Eric Tepe
> ___________________
> Biological Sciences
> University of Cincinnati
>
>
>
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