Apramycin plates - (Nov/19/2005 )

When I transform my E. Coli cells with apramycin resistant plasmids and plate them on Apr plates, they grow just fine. When I pick colonies and inoculate them into liquid media with apramycin they do not grow... What is going on? Something must be happening to my cells between plating and inoculation... could putting the plates in the fridge possibly be lowering or eliminating the viability of the cells?

Dave.

-zidave-

Do your plates and the liquid medium have the same apramycin concentration?
The same used to happen to my transformants with ampicillin due to a too low concentration
of the antibiotic in the plates...

-speedy-

They are the same concentration. There must be something else happening.

-zidave-

I am assuming you add your antibiotic to cooled media, and all the obvious stuff like that?

and how long are we talking, in the fridge prior to subculturing?

-aimikins-

QUOTE (aimikins @ Nov 20 2005, 07:03 PM)

I am assuming you add your antibiotic to cooled media, and all the obvious stuff like that?

and how long are we talking, in the fridge prior to subculturing?

Less than a week.

-zidave-