Freeze and Squeeze

[Robert Horton]

Kevin Simcox (simcox.3 at osu.edu) wrote:
: > Ann M. Yezerski wrote:
: > >
: > > Does anyone have the standard protocol for "Freeze and Squeeze" to remove
: > > DNA from an agarose gel?  If so, e-mail me at ayezersk at moose.uvm.edu.
: > 

: Take a 500 ul microfuge tube and make a small puncture in the bottom 
: using a small gauge syringe needle.  Place a small amount of 
: silconized glass wool in the bottom.  Cut out the band of interest and 
: place into the 500 ul microfuge tube.  Then place the 500 ul microfuge 
: tube into a 1.5 ml microfuge tube.  Freeze at -70 C for 20 min.  Spin for 
: 10 min and collect liquid containing DNA fragment. The DNA fragment can 
: be equilibrated in a 0.3 M NaOAc, 0.01 M MgCl2 solution prior to loading 
: the microfuge tube.  Just add 2.5 vol of ethanol after centrifugation to 
: ppt the DNA fragment.

REFERENCE:
Tautz, D. and Renz, M. (1983) Analytical Biochemistry 132:14.


---
Robert M. Horton
http://134.84.47.3                                       Have a :) day

"Scotty, try flushing the radioactive waste into the ventilation system!"