fly DNA extraction
5764ottoj at vms.csd.mu.edu
Fri Sep 16 10:17:40 EST 1994
In article <1994Sep16.203710 at ccvax>, mbyhsun at ccvax writes:
>What is the best way to isolate DNA from Drosophila? I need to get DNA for
>southern blots. Thanks.
I have had good luck chilling them in a mortar and pestle to dry ice temp,
then grinding them up. I then transfer the fly powder to a homogenization
buffer (unfortanately don't have the composition here), vortex and pellet
the nuclei. Wash a few times in an extraction buffer, add proteinase K and
SDS to 100ug/ml and 1% respectively. Cook for 1h at 50C, Phenol extract,
til no visible interphase, Phenol:Chloroform extract 3X, Chloroform extract
3X, add NaCl to 200mM and then add 2.5 vol 100% EtOH. I then spoon out
the genomic DNA with a glass hook, and wash several times with 70% EtOH.
If this method sounds good to you, email me back and I will supply you
with all the particulars.
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