salmon sperm DNA

[Marieke R. Koedood Zhao]

We don't extract either - we dissolve the DNA, shear it by pulling through a
syringe needle multiple times and then boil it for 10 min. That's it.
Works fine for hybridizations as well as yeast transfections.

Marieke


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