supF plasmid libraries and growth of MC1061/p3 or DH10B/p3 bacteria

Mark Worthington mtw3p at AVERY.MED.VIRGINIA.EDU
Sun Nov 22 13:02:54 EST 1998

I have received libraries in pCDM8, a supF containing plasmid through which
antibiotic selection is mediated through complementation of an amber mutant
in the AMP and TET resistance genes on the p3 episome.  In two strains
containing this episome (MC1061/p3 and TOP10/p3), the colonies come up
fairly late (some at 16 hours, more at 36 hours) and, as a result, there is
a range of colony sizes.  The concentration of Amp and Tet in the plate
makes no difference, as far as I can tell.

Is this a normal feature of this plasmid?  Any thoughts about when to
harvest the plates?

Thanks in advance for your assistance.

Mark Worthington
University of Virginia

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