how to ligate two ends with different enzyme site - (Jun/17/2006 )

I want to delete a short fragment from a vector. So, have to use Bgl II and BamH1 to cut it out and then ligate the linearized vector together. However, the 3'end is BglII and 5'-end is BamH1. How to liagte the two ends?

any advice will be appreciated highly.

Thanks

Lifan

In this cases one should do either a fill in or an exonuclease digestion, in order to obtain blunt ends and then you can clone it. This unless the two ends are compatible (you can check in the last pages of the catalogues of enzymes selling companies), in which case you can ligate directly. If this piece is in a coding region, check the frame.

Hi Lifan,

I think you can ligate the cut out vector (5' BamHI and 3' BglII). This is because they are compatible. I have experience with this. The only thing you should know is that the fusion site will neither be cut by BamHI nor BglII. It is because the site becomes semi-BamHI and semi-BglII.

Cheers,

Charles