blunt ligation problem
Patrick F.H. Lai
pfhlai at LOOKSMART.COM
Tue Aug 18 20:10:41 EST 1998
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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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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 >
University of Toronto
Toronto, Ontario, Canada
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