Freeze and Squeeze

[Kevin Simcox]

> 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.

Kevin Simcox
simcox.3 at osu.edu