Temp. for DNA/glasswool binding?

Ed Beaty ed_beaty at qms1.life.uiuc.edu
Mon Apr 25 15:49:33 EST 1994


Hello, hello again...

I've been trying to use saturated NaI to dissolve agarose gel slices and
bind the DNA trapped therein onto acid-washed glasswool (which is easier to
handle than the various loose Celite/diatomaceous earth particles I've
tried).  I get some (30% ?) recovery of DNA after washing the glasswool,
drying it, and eluting the DNA in TE buffer at about 60 C.  Of course, I'd
like something closer to 100% recovery...

I'm thinking about raising the temperature of my NaI/DNA/glasswool mixture,
to increase the rate of DNA binding.  However, I'm not sure whether raising
the temperature to 60 C or so will increase the rate of binding, or will
have the opposite effect.  Has anyone out there tried this?  Or any
educated guesses about how temperature will affect binding?  And is
irreversible binding of the DNA to the glasswool a problem at higher
temperatures?  

Thank you!

Ed Beaty
Ed_Beaty at qms1.life.uiuc.edu



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