5' protruding ends

Dom Spinella dspinella at chugaibio.com
Fri Oct 3 19:04:14 EST 1997


> 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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