pkursula at cc.oulu.fi
Wed Mar 5 05:51:03 EST 1997
On Tue, 4 Mar 1997, T.J. Young wrote:
> 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?
> 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.
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