Transformation with ADE2

Stephen Cronin scronin at jeeves.ucsd.edu
Mon Jun 2 18:40:41 EST 1997


Does anyone have experience with integrating vectors using ADE2 as
a selectable marker for transforming ade2-101 strains?

   When I transform I get a small number of large white colonies plus
about ten times as many very small colonies that never reach full size.
When I pick these small colonies and streak them out onto fresh plates,
I get a mix of large and small colonies.  The vectors I am using are
based on pASZ11 and pASZ10 (Stotz and Linder, (1990)GENE 95:91-98). I
have been using a Lithium acetate transformation protocol and selecting
on minimal media with the appropriate supplements.  Have you seen this
transformation phenotype before, or something similar?

Thanks,
Stephen Cronin
scronin at jeeves.ucsd.edu



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