Phosphatase Columns

Andrew G. McArthur AMCARTHU at UVVM.UVIC.CA
Fri Mar 3 15:03:44 EST 1995


Colleagues,

We have been making a TA vector out of M13mp18 for some time now by
using HincII and Taq addition of a single T overhang.  However, we have
been having troubles with recirculization of the molecule with no insert
during ligation.  The single T leads to a frameshift mutation in lacZ and
thus a white plaque.  We wish to dephosphorylate our TA M13mp18 prior to
ligation and are curious about the BIO/CAN Scientific Alkaline Phosphatase
Compact Reaction Columns.  Does anybody have experience with these columns -
are they a good investment?  Does anyone else who uses home-made TA vectors
have an in-house protocol for dephosphorylation (it is not something we
have had need to do before)?  Much thanks for your help, as usual.

Andrew McArthur
The Koop Molecular Evolution Group
University of Victoria
Canada
amcarthu at uvvm.uvic.ca



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