Best method for DNA extraction from fixed tissue

beckwith at netcomuk.co.uk beckwith at netcomuk.co.uk
Mon Apr 19 15:30:40 EST 1999


GR Taylor wrote:
> 
> Does anybody have a really good method for getting DNA out of fixed
> pathological tissue, either commercial or Home Brew?
> 
> Our experience has been that shorter times of fixation have given better
> yields, and that extended exposure to proteinase K improved efficiency.
>  Salt extraction or phenol seem to work about as well as each other.
> 
> We have tried several approaches, most work to some extent but fragments
> larger than 200 bases get recovered with low efficiency.  Getting larger
> fragments out would be very useful.
> 
> Perhaps there some organic chemists reading this who can tell me what
> formaldehyde does to DNA and protein during the fixation, and if there is
> anyway of reversing any of the effects.
> 
> Graham Taylor
> DNA Laboratory,
> St James's University Hospital,
> Leeds
> UK
> http://www.leedsdna.demon.co.uk
> tel (44) 113 206 5217
> fax (44) 113 246 7090


Ex-Wax DNA Extraction kit for paraffin-embedded tissue.
beckwith at netcomuk.co.uk,for further information, if interested, as not
to advertise the company!



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