insert-positive blue cols

Fiberman wsun at jeeves.ucsd.edu
Sat Jan 23 18:15:35 EST 1993


In article <1993Jan21.152404.26275 at magnus.acs.ohio-state.edu> cbreiten at magnus.acs.ohio-state.edu (Caroline A Breitenberger) writes:

[stuff deleted]
>
>In another case, a graduate student who is using the Clontech TA cloning
>kit (:-)) noted that clones containing his desired PCR-derived insert
>(appr. 250 bp) exhibit a pale blue phenotype.  In this case, I think
>that the insert provides a continuous open reading frame which results
>in expression of the alpha-complementation peptide with an N-terminal
>extension.  The pale blue color suggests that this peptide is less
>efficient than that of the parental vector.  (The opposite orientation
>of the insert presumably gives white colonies - I forgot to ask him.)
>
>Caroline Breitenberger, Department of Biochemistry
>Ohio State University, 484 W. 12th Ave., Columbus, OH 43210, USA
>614-292-9473.......caroline+ at osu.edu


I have had the same experience.  I tried calling Stratagene,
asking them where does the alpha complemetation peptide start in
the bluescript vector and how long is the peptide, but the
technician did not know anything.  If the peptide is short
enough, cloning an insert into a site that's at the 3' end of
the polylinker will not give you any white colonies.  Any one
out there knows how long is the alpha comlementation peptide?

-fm



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