Blunt-end troubles

Colin Rasmussen crasmussen at
Wed Mar 12 20:56:12 EST 1997

In article <33254473.49E4 at>,
bgerhar at wrote:

> I had a similar experience with a sticky end ligation where I wanted
> both orientations of a BamHI insert. In this situation, I could never
> get the opposite orientation. I was using blue/white selection and found
> that when I didn't use IPTG/Xgal I was able to get the second
> orientation. I am not sure if this is something that you are doing, but
> maybe it will help.

It probably means that in one orientation, expression of your insert (for
whatever reason) was toxic to the cells.

> Good luck,
> Brenda

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