colony hybs

Martin Kennedy mkennedy at chmeds.ac.nz
Sun Sep 19 23:01:37 EST 1993


In article <leach-170993114747 at med-pharm5.bu.edu>, leach at mbcrr.harvard.edu (Martin Leach) writes:
 
> Hi,
> I have a simple procedure.
> I use gridded Hybond N nylon membranes from AMersham.
> Place the circular membrane on the bugs for 3'
> mark membrane, then place colony side up for 3' on 3MM Paper soaked in
> denaturing solution,
> then 3' on neutralizing solution soaked 3MM paper
> then 3' on 2xSSC soaked paper.
> I then use the Stratalinker UV-crosslinker to stick the DNA to the blots.
> Alternatively, you could use a vacuum oven to bake the blots.

I use an even easier method, "turbo lifting" by Buluwela et al.
( NAR 17, p452).  Do your lifts onto filters, place them onto 3MM
paper soaked in "turbo juice" (2xSSC, 5%SDS), and microwave the 
whole lot for 2.5 minutes.  You can optionally UV cross-link
but its not necessary. 

Cheers,

Martin

NNNN   NN  Martin A Kennedy (E-mail = mkennedy at chmeds.ac.nz)  ZZZZZZZ  
NN NN  NN       Cytogenetic and Molecular Oncology Unit          ZZZ
NN  NN NN           Christchurch School of Medicine            ZZZ
NN   NNNN              Christchurch, New Zealand              ZZZZZZZ
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