blunt ligation problem

[Patrick F.H. Lai]

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In answer to queries:  1.  To generate a blunt ended plasmid, I cut it with SmaI.  2.  I did also dephosphatase the plasmid.  This worked as I tested this by comparing normal digested plasmid to dephosphatased plasmid when transforming the bacteria.



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HI !

Do you plan to keep the SmaI site ?
If not, how about adding 3'-overhang A's & T's to your inserts and
vectors and perform TA-ligation ?
Just a thought.....






Hope this is useful info.   :-)


Patrick F.H. Lai  < PFHLai at looksmart.com >
Graduate Student
University of Toronto
Toronto, Ontario, Canada





       
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