leach at bu.edu
Mon Jan 15 16:23:45 EST 1996
check out the Jan 96 issue of biotechniques...
there is a nice article on increasing the efficiency of T-vector ligations...
basically in the preparation...
1. digest the vector with a blunt cutter (we use EcorV)
2. do the T'ing reaction using Taq pol
3. Set up a ligation reaction...self-ligating the t'ing reaction
any non-t'd vector will ligate to other non-t'd vector forming either
circular plasmids or concatamers of it.
4. Run the ligation reaction on the gel and excise the linear form...that with
a T at each end of the vector.
....increases the efficacy of cloning since the t-vector prep is now enriched
for the correct construct.
..... Martin Leach Email:leach at bu.edu
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