salmon sperm DNA

J. Pat Martinez martinez at puccini.CRL.umn.edu
Sat Dec 21 06:08:10 EST 1996


We dissolve it (from Sigma) in water then autoclave the solution for 15 -
20 minutes to shear it (much easier than using a sonicator).  We don't
bother to extract it with organics.  Works fine in our prehyb. soln.

Pat M.

In article <32B7F7AD.43BC at anatomy.anat.med.ualberta.ca>, darren cikaluk
<dcikaluk at anatomy.anat.med.ualberta.ca> wrote:

> Does anyone know how to properly prepare salmon sperm DNA?
> 
> Manniatus suggests to dissolve it to 10mg/ml, shear it, then extract 
> it with phenol/chloroform.  Shearing it is no problem but i tried to 
> extract it and it just looked like glue. After spinning it to seperate 
> the phases, the aqueous layer looked terrribleand very cloudy.  I'm 
> sure it is way too viscous and too conc'd to extract.  Therefore can 
> you get by without extracting? or what do I need to do.
> 
> Ps we will be using this for cell culture transfections and as a 
> blocking agent for blots.
> 
> darren



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