Plant DNA extraction - ethanol and dried plants

[Txn]

For problematic tissues (ironically I have more problems often with
fresh tissue than freeze-dried but of course most herbaria tissue is not
freeze-dried) i use a derivative of a method from Warude et al (2003)
Plant Mol. Biol. Rep. 21: 467a-f. It is basically a detergent based
method with alcohol precipitation. The main changes you can try are to
increase the amount of detergent used (if you suspect the cells are not
being permeabilized properly). They suggest checking the pH after the
initial extraction (prior to alcohol ppt) since plant tissues are often
pretty acidic - in this case you can just add more buffer. Addition of
polyvinylpyrrolidinone (PVP) or polyvinylpolypyrrolidinone (PVPP) or
beta-mercaptoethanol at higher concentractions can help if you suspect
tannins and polyphenolics are a problem. Lastly in this paper they
increased the NaCl concentration to help with polysaccharides problems
(often noticeable i find as a gel like-DNA product.). Hope that helps

-Ben


Vitor Miranda wrote:

> Dear colleagues,
>
> I have extracted DNA from fresh plant material with no problems, but
> from dried (herbaria) tissue it has been a real problem.
> Does anybody know a successfully protocol to extract DNA from herbaria
> material?
>
> Other question: I never tried to use material preserved in ethanol. Is
> it possible to get DNA from this kind of  material?
> Any suggestion is very welcome ;-)
>
> Thanks in advance,
> Vitor.
>
> --
> --
> Vitor Fernandes Oliveira de Miranda
> Professor Assistente
> Universidade de Mogi da Cruzes - UMC
> Laboratório de Sistemática Vegetal
> Av. Dr. Cândido Xavier de Almeida Souza, n.200
> CEP 08780-911 - Mogi das Cruzes - SP - Brasil
> Tel.: +55 (11) 4798-7143/ 4798-7313
> http://www.umc.br/~vmiranda/lsv
> e-mail: vmiranda at umc.br