freeze-squeeze

Michael Gillings gillinm at agric.nsw.gov.au
Sun Mar 3 17:58:45 EST 1996


I have used a method from low melt agarose that we used to call the 
"Freeze-squeeze". Cut your band from the agarose and place it into a 
folded piece of parafilm in the freezer.  When it is completely frozen 
place the agarose parafilm sandwich between two sterile microscope 
slides. Make sure that some of the parafilm sticks out the sides of the 
setup.  Form a "V" with the microscope slides that is in the same 
orientation as the "V" of the parafilm.  Slowly squeeze the slides 
together, and as the slice defrosts under the pressure, the buffer and 
some of the DNA will come out of the agarose. Collect the droplet of 
buffer from the parafilm fold.
This works best for small molecular weight fragments (naturally).  Phenol 
extraction will remove the bromophenol blue.

Good luck,
Michael


Dr Michael Gillings
Senior Research Scientist
Biological and Chemical Research Institute
PMB 10, Post Office Rydalmere
NSW 2116
AUSTRALIA

Ph   61-2-843-5709
Fax  61-2-630-4475
On Thu, 29 Feb 1996 vsood at acs.ucalgary.ca wrote:

> I remember seeing some advice on retrieving DNA from agarose slices via a
> method of freezing the slices and then mashing them, but don't recall the
> details. Does anyone use this method who can refresh my memory? Also, what
> about any EtBr or bromophenol blue (my band runs at the same place as the
> dye front) present in the slice? 
> 
> thanks a lot. 
> bb.
> 
> 



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