epitope duplication

rogier stuger at cellbiology.uni-frankfurt.de
Wed Jun 30 05:22:35 EST 1999


Hi,
how big is your epitope?
If it is small enough, just stick some oligos in your
plasmid.
If it's too big for oligos, PCR it out with overhanging
primers for IDENTICAL restriction sites, ligate the (cut)
PCR product into your vector at a high insert-to-vector
ratio (try 25:1).
After transformation, check your clones for # of inserts by
restriction of minipreps or by PCR on your clones (can be
done directly on E. colis) using primers just outside your
epitope insert

Good luck,
Rogier, MicFizz, Free U of A



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