Genomic DNA extraction
jrineharNOSPAM at snake.eou.edu
Tue Jul 30 12:19:51 EST 2002
For several years now we have been using the Promega Wizard genomic DNA kit.
This works with almost any insect (we've used it successfully with flies,
even single flies, where we get nearly a microgram of DNA per fly; we've
also used it with more "crunchy" insects like bark beetles). The trick is to
make sure the animals are ground but not macerated, because that seems to
cause shearing. Otherwise, it's a very good little kit in our experience.
Hope this helps.
Andrea Osterlehner wrote:
> I am struggling with the isolation of genomic DNA of Drosophila. First I
> tried a tissue kit which worked well for marine organisms but did not
> work for Drosophila. Next I tried a protocol using frozen flies,
> grinding them in a buffer containing SDS, Sucrose and EDTA, heating,
> Phenol-Chloroform extraction and final EtOH precipitation. The DNA yield
> was poor too, only 140 ng from 15 flies. For a southern blot experiment
> I need much more genomic DNA.
> I would be glad if somebody could send me a protocol, which works well.
> Thanks a lot, Andrea.
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