Blunt-end troubles

Andrew Doherty A.Doherty at
Wed Mar 5 11:47:12 EST 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 ;-)
The only thing I can think of is that the insert is lethal when it's the
right way round - other than that, I'm buggered if I know!!!!
Hope it helps

Andy D 
Dr Andrew Doherty		email -  a.doherty at
Dept. Anatomy			Tel (0117)9287421
School of Medical Sciences	Fax (0117)9287402
University of Bristol
University Walk
Bristol UK

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