Blunt-end troubles

Colin Rasmussen crasmussen at anat.med.ualberta.ca
Wed Mar 5 11:10:36 EST 1997


In article <Pine.SOL.3.91.970305124952.26779A-100000 at sun1.oulu.fi>, Pete
<pkursula at cc.oulu.fi> wrote:

> On Tue, 4 Mar 1997, T.J. Young wrote:
> 
> > Hi,
> > I've got a bit of a problem, I'm trying to subclone a DNA sequence from one 
> > vector into another.  Unfortunatly, the MCS are not compatible so I'm using 
> > blunt-ended ligation.  Now this is a bugger at the best of times but every 
> > time I get some positive colonies, on inspection, I find that the sequence 
> > has gone in the wrong way round!!  Now am I just the Worlds unluckiest 
> > molecular biology student or could there be some kind of underlying 
> > phenomenon that's driving my ligation in this direction?
> > Cheers,
> > Tim ;-)
> > 
> Yes, it is a general phenomenon: your insert always goes in in the wrong 
> orientation, if it is possible. 
> 
> ;-) At least that is my experience.

I think there's a Law by Murphy that cover this one.

Colin Rasmussen



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