PFGE Southern Blots

Antje Haase antje at menzies.su.edu.au
Wed Aug 3 21:39:45 EST 1994


> 
> Does anyone have any tips regarding transfer of large DNA fragments
> (50-250 kb) to nitrocellulose or nylon membranes?  Is depurination 
> necessary?  How many times can the membranes be stripped and reprobed?
> 

Hi Walter,

we routinely UV our PFGE's on a short-wave trans-illuminator for a couple of
minutes and then do the regular denaturation and neutralisation for
Southerns. It works fine for even larger than 500 kb fragments. 

As far as stripping and reprobing is concerned this will depend on the
membrane you are using but is shouldn't be different to any other blot.

Hope this helps

Antje

_____________________________________
Antje Haase
Menzies School of Health Research
Australia
______________________________________
> 







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