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
More information about the Methods
mailing list