Preparartion of Liver DNA for PFGE

Dr Saleem Mohammed saleem at
Wed Mar 8 23:10:22 EST 1995

I was wondering if anybody had  a method  for analysing rat liver genomic DNA 
by pulse field gel electrophoresis.  I have talked to a number of people who 
suggest using agarose plugs.  

The question is how does one break liver cells from frozen liver (liquid 
nitrogen).  Could one just homogenise using a pestle and mortar and then 
prepare agarose plugs from the powdered liver and if so how much liver to use 
per sample. 

 The alternative is to isolate genomic DNA using traditional 
proteinase K, RNAase and phenol/chloroform extractions, although this can 
damage genomic DNA.

ANY IDEAS would be greatly appreciated.

Many thanks

Dr Saleem Mohammed,
Dept of Biochemistry & Molecular Biology,
Royal Free Hospital School of Medicine.
Rowland Hill Street,
London. NW2 2PF.

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