5' protruding ends

Geoffrey Kidd GKidd at Aptagen.com
Tue Oct 7 15:32:57 EST 1997


I second what Dom wrote.  

Geoffrey Kidd

Dom Spinella wrote:
> 
> > Hello
> >
> > Just wanted a quick opinion regarding converting protruding termini
> > to blunt ends from those out there who do this more regularly than I.
> > I have used Mung-bean nuclease for this in the past, but perhaps there
> > is something better available. Also, for my general knowledge, I recall
> > that Mung-bean is often considered a better choice for this than
> > nuclease S1 but don't remember the exact reason.
> >
> > Thanks for your help,
> >
> > Craig Garen
> 
> Craig:
> Actually, I think most folks just fill in 5' overhangs with Klenow or T4
> polymerase and dNTPs.  These enzymes also have 3'->5' exonuclease
> activity and can therefore also be used to remove 3' overhangs. Mung
> bean and especially S1 nuclease can be a little "agressive" and
> sometimes chew back even the double stranded DNA, so a lot of people
> don't use them for this purpose anymore.  Hope this helps.  -- Dom
> Spinella



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