Hi Andreas,
if the kanamycin degrading enzymes were the remains of a one year old (dead) sample without newly produced enzyme, they should be degraded by other proteases and been long gone. Although unlikely, a tiny bit of the enzyme might be still intact but not in a quantity required to clean your LB medium.2015-01-26 11:54 GMT+01:00 Mega [Andreas Stuermer] <masterstorm123@gmail.com>:
Hi guys,--
For a workshop I gotta revive agrobacterium + GFP from the fridge.
I still have a one year old petri dish and a liquid culture, and a stab culture. Strangely, I got no colonies from the stab culture on a petri dish yet (1 day, but I know they grow very slow). Should have done fresh stabs half a year ago it seems. I always thought stab cultures are most durable?
I also tried putting the liquid culture on a petri dish, which also yielded no colonies after 1 day.
So finally I tried to pick tons of bacterial lawn/colonies from the petri dish, put them in liquid LB kan medium and it got a bit cloudy after one day.
Maybe the explanation - in liqiud medium the end-products are distributed more homogenously - so quicker growth but quicker stagnation/dying.
I was wondering, do the kanamycin degrading enzymes still work after one year?
Would you assume it's still the correct bacterium after one year - contamination wouldn't grow as quickly I believe. (I could do a transient GFP plant, but it takes 5 days and I would have to send it today)
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