Andrew Bradbury bradbury at ICGEB.TRIESTE.IT
Fri Apr 25 10:30:44 EST 1997

Sounds too easy, but I'm having problems with changing the ampicillin
resistance of a pUC119 based plasmid with any other resistance.  I am
cutting with BspH1, which flanks lactamase, and trying to clone in a PCR
fragment from Alexeyev's cassette antibiotic resistances (chloramphenicol,
gentamycin or kanamycin).  I get resistant clones, but they are all
rearranged.  Is there something special about the bit between the two BspH1
sites?  Any suggestions? Am I doing something silly?
I have been thinking of amplifying the plasmid, excluding amp, and just
adding sites to the end of the PCR product.  Any suggestions about which
bits I should keep and which I should not?

thanks for your help

andrew bradbury

Andrew Bradbury

Area di Ricerca
Padriciano 99
Trieste 34012

Tel +39 40 398995
Fax +39 40 398991

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