T-Vector for PCR cloning
findley at u.washington.edu
Tue Feb 1 11:58:25 EST 1994
I have seen a few postings recently regarding T-vectors.
I have one almost made, but have little interest in finishing the
construction. If somebody wants to finish this, I can send the stuff:
I cloned two T end-producing restriction sites (Eam1105I) into the polylinker
of pBS.(There were very few choices for how to do this).*
There is unfortunately still another Eam1105I site in pBS ampicillin(R) gene
that still needs to be converted.
I have made an oligo to delete the site via a silent substitution (by any
number of techniques).
If somebody wants to do the mutagenesis, I can send you: the oligo, the
T-site polylinker and a cheery "good luck to you!"
* I used an couple of oligos to introduce a small fragment into pBS-II-SK
polylinker. The fragment (20 bp) contains two Eam 1105I sites (NEB) in an
inverted orientation relative to one another.(look up the site).
Because the recognition site has a few internal N's I made them produce
ends which have the required T overhangs.
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