pUC18/19 double digest with BamHI/SmaI

bweil at mzdmza.zdv.uni-mainz.de bweil at mzdmza.zdv.uni-mainz.de
Thu Jul 13 02:26:25 EST 1995


In article <3s4ahn$nka at hippo.shef.ac.uk>, M.C.Behrendt at shef.ac.uk
(M.C.Behrendt) wrote:

> Has anyone tried to do a double digest using BamHI and SmaI with pUC18 
> or 19.  I'm just wondering because the two sites on the MCS overlap by
> one base, will this cause problems?
> 
> Mark


Hello,

You can take a puc-vector with an BamHI Insert in it.
Then you can cut with SmaI (in the MCS) and then with BamHI. This should
cut the distant BamHI-site in the MCS. If the Bam-site near the Sma-site
is cut does not matter. You can then separate Insert and Bam-Sma-cut
vector and insert by electrophoresis and elute the vector.

good luck,

Bernd Weil



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