slimy pellet - (Oct/24/2007 )
I[size="3"] am working on a recombinant e. coli. i am facing a problem which is really haunting me. after centrifuging the broth the pellt i get is very slimy....and it becomes very difficult to prepare a homogenous slurry so to be able to proceed for the disruption........can anybody throw some light...????
a slimy pellet?
how many ml of medium are you centrifuging, what's the speed and time of centrifugation?
are you certain your culture has not been contaminated? E coli isn't particularly slimmy. Does the colour and smell look alright?
If you have a large paste, remove the supernatant and vortex the pellet. You will find that the pellet breaks up when vortexed in this manner.
If you have a large paste, remove the supernatant and vortex the pellet. You will find that the pellet breaks up when vortexed in this manner.
even my doubt is for the contamination...i dont get the meaning of your last line
The last line was just incase you are facing a larger then normal pellet. Big pellet can be sticky and don't suspend easily.
In any case, best to grow the culture again, fresh. Preferably with a different isolate of your desired strain.
Slimy apprearance sounds like your bacteria are half dead before your harvest.
and why does that happens?
parveen, if you can you might like to have someone experienced look at your pellet next time.
I would describe e coli as pretty slimy. on the other hand, lysed cells are *really* slimy. and, very tedious to resuspend.