bjornrosen at swipnet.se
Wed Aug 13 15:04:25 EST 1997
Helene Bach wrote:
> Does anyone how to isolate DNA from liver? (My preps have alot off
having worked in a forensic laboratory, you can imagine what kinds of
tissues I have been extracting DNA from. And they were not always in a
very good state (to say the least).
Thus, I have extracted DNA from liver and with succes. The protocols I
used were either standard phenol-chlorophorm extraction or a
Chelex-based quick-prep method (if you would like to receive protocols
or references, let me know).
I guess your problem is the same as I used to have; inhibitors of the
DNA polymerase in your DNA extracts. One easy way to remove inhibitors
is to use Microcon or Centricon micrconcentrators. They can be used to
1) concentrate the DNA extract, 2) wash the DNA extract, 3) change the
solution in which the DNA is dissolved. These microconcetrators are
manufacured by a company called Grace (no association). Since you can
use the microconcentrators to wash the DNA, you can use any solution
(water, buffer with high/low salt and with or without detergents) for
the washes, and then the final wash would be in water.
The Microcons are the better choice because they fit in any
microcentrifuge for 1.5 mL Eppendorf tubes. My experience of other
brands than the concentrators from Grace is not very good. With Microcon
and Centricon the recovery is close to 100 per cent. I should maybe
stress that I don't work for Grace once again, but I do think they have
a great product.
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