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5-FOA

Martin Hirst hirst at interomex.com
Tue Apr 22 11:30:26 EST 2003


Are you sure that your plates are correct - how are you making them?  Try
patching a ura+ and ura- strain onto the your plates to be sure they are ok.
Generally we plate 10-100,000 cfu's on a large plate (20cm).

Cheers

Martin

> Have been trying to use 5FOA to dedelect for URA+ to no avail. I've
> tried adding proline to the media in order to make the cells
> hypersensitive to 5FOA and have also tried replica plating to new 5FOA
> plates after initial growth.
> 
> The problem is that after each try, instead of only a few cells
> growing, I find a lawn of cells. How many cfu/ml should I be spreading
> in the first place, and also is there anything I might be missing?
> 

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