How do you extract fungual/bacterial genomic DNA? - (Mar/20/2006 )

HI!
I try to extract bacterial AND fungual DNA form leaves and noted few minutes ago that the protocol i was using is futile - I got DNA, but the extraction was not very effective - and maybe it was selective. Whatever - what are your "personal" protocols for extraction?

CU

Sven

-dersven-

Maybe you could try to extract bacteria first, and then fungi with 2 different protocols?

-yata-

Do you first culture the bacteria and fungi?
Cause you need a lot wet weight of fungi to get a good amount of DNA because you have to powder them.

-aspergillie-

QUOTE (aspergillie @ Mar 22 2006, 05:08 PM)

Do you first culture the bacteria and fungi?

I don't culture the bacteria.

What are your protocols for extracting (cultured or not,...)

-dersven-

To extract DNA from a fungus I let them grow o/n in complete (rich) medium. Then collect the mycelium by pouring the culture through "miracloth". Grind the mycelium in liquid nitrogen. Add a spoon of the powder in Extraction buffer (400µl). Twice a phenol-chloroform extraction, ethanol precipitation (here without washing with 70%), RNase treatment, additional phenol-chloroform extraction and a complete ethanol precipitation. Then the DNA was really clean and nice.
But it's not clear to me if you want to extract DNA from a bacteria/fungus mix or seperate.

-aspergillie-