help on blotting megabase DNA

Nick Jacobsen jacobsen at cf.ac.uk
Thu Jul 3 06:03:46 EST 1997


Hi,
  For PFGE blots I depurinate the DNA by soaking the gel in 0.25M HCl for 
12 minutes prior to denaturation/neutralisation and transfer. I have had 
intermitent trouble with Amersham Hybond N and N+ and as a consequence I 
use Bio-Rad Zetaprobe GT nylon membrane which sticks DNA like shit sticks 
to a blanket if you pardon the analogy!!! I have reprobed my PFGE blots 
upto 20 times when using the above methods


hope this helps


Nick.




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