I am working with a bacterium which is very slimy so I am not able to get the tight pellet, but i need to get tight pellet with very less water for further analysis(pigment analysis)...even i tried to centrifuge at 14000rpm for 30-50 minutes still no use.... Please help me.
Amylase in saliva digests certain polysaccharides. Research and experiment along this line of thought. Must the bacteria remain viable? For what are you assaying?
Dennis R. Hill
I am checking for the effect of stress on pigmentation, once the bacterium reached stationary phase I will stop bacterial culture at different time point. After incubation I need to get pellet (I will extract pigment using acetone)then I will analyze the pigment. As I am not getting the tight pellet and which contains the water inferring with my result.
I cannot dry the pellet and do my work, please suggest me some other alternatives.
Thank you.
not really sure how your assay works... but perhaps you can consider designing a control (eg stress-free bacteria) and use it to offset the effects of water on your assay?
Most of the time, this "slime" is from exo-polysaccharide that is produce by the organisms. You will have to wash the cell with a saline physiological solution (0.85% NaCl) or a similar physiological solution. Pellet the cell at low speed by centrifugation. Resuspend them in a similar volume by the saline solution (same volume as initial working volume). Wash again one or two time using the same technique. Your cells should be ready.
Try with "sputasol" and physiological solution (0.85% NaCl) eluited with amylase, try for 3 times whashing and centrifugating process, it should work.
What kind of bacteria is it?
Although not to much effect…try using centrifuge tubes with conical bottom (Falcon tubes for e.g.) against round bottom tubes (oakridge). The conical shape helps to hold the pellet more tightly…...
Ok, so you could do what I did with a P.aeruginosa some weeks ago, that was very slimy.
Add the "sputasol" directly into a Falcon (15ml or smaller if you find) with 3/4 colonies of your Chryseobacterium, vortex a lot, then centrifugate. Use the deposit to make another sospension with physiological solution and amylase. Vortex, centriifugate 3000rpm/5min and then if is still not enough, repeat.
Whit my Pse.aer it worked!
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