5 prime overhang removal

Peter Ashby p.r.ashby at dundee.MAPS.ac.uk
Wed Mar 29 03:19:12 EST 2000


 mclean at NOSPAMaecom.yu.edu (Gary McLean) wrote:


> mlsulliv at facstaff.wisc.edu ("Michael L. Sullivan") wrote:



> > >I need to chew back a 5 prime overhang to produce a blunt end. According
> > >to the Promega catalogue I could use either Mung bean nuclease or S1
> > >nuclease. I am familiar with using the Mung bean for 3prime overhang
> > >removal but I'm not sure about 5 prime. Can anybody comment which would be
> > >best or recommend something else?
> > 
> > I've used mung bean nuclease a couple of times for blunting fragments with
> > 5' overhangs, and it worked just fine.  I just followed the protocol that
> > NEB has with their enzyme.
> > 
> > Mike


>  I've also used the NEB mung bean with mixed results...I would suggest
> following their protocol but make sure there is a lot of dna present and
> the mung bean is freshly purchased. This enzyme can digest away dna like
> crazy!

Thanks for the info everyone, I will try it. I made it work a while ago
for 3prime removal so am mildly optimistic.
 
> G
> 
> -- 
> Lets go Otago!!

Seconded

-- 
Peter Ashby
Wellcome Trust Biocentre
University of Dundee
Dundee, Scotland
Reverse the spam and remove to email me.




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