What I have:
I've cultured certain becteria by mixing bacterial suspension with agar cultural media before solidification. Now the bectria cells are dispersed inside the agar gel.
What I want to do:
In order to conduct metabolomics experiment about the physiology response of the becteria grown in solid matrix versus liquid condition, I need to collect the bacteria cells from the gel.
What I want to avoid:
1. the destruction of becteria cells.
2. large amount of substances in matrix mixed with bacteria after separatio.
3. poor repeatability and poor yiel.
Can anyone give me a help?
hi
if the bacteria still in agar you can take one or two colony of bacteria from agar by sterilized loop and culture in broth media like nutrient broth and incubate it in shaker incubator.
and if you have a stock of bacteria you can culture the bacteria on the surface of nutrient agar not mixing with it or in broth media.
good luck
Just pick one single colony and subculture your agar(solid) or broth (liquid) and put them into incubators. If you want to froze your bacteria, make sure to use 15% glycerol and pick all these microbes into the solution then store them at -80°C so that you can recover your bacteria anytime.
Hi there,
Not clear if it's a mixed bacterial culture.
Better you culture separately on agar surface with single strain.
@ Chen Shanqiao: Why do you not culture bacteria on agar surface? It is growth on solid media. If are you interested in oxygen limited growth, you can use surface growth also, you need only to control growth atmosphere.
Regards
Vit
Hi Chen Shanqiao,
Not clear if your bacteria require an anaerobic or miro anaerobic culture atmosphere. Try to culture it on agar surface, or if it is micro aerophilic, then wrap the culture plate with several rounds of parafilm to limit O2 access. Good luck.
Hi brother,
I think the method of separating bacteria from the solid media depends on the temperature of the bacteria.
Your samples are inside solid media, are not you?
Is it possible to find one colony on the surface of the media?
If there is no colony, the media must be heated according to the temperature of the bacteria until it gets liquid, then take one ml of the media and cultured on a nutrient agar. then The plate incubated for 24 h at 36ºC ± 1ºC,
first ,separate the colonies Required by loop .second, save it in 16% (v/v) glycerol in brain heart infusion broth (Oxoid Ltd, UK) and were stored at -70˚C(Rowan et al., 2008)
Regards .
Hi @Chen Shanqiao, I hope you already got the answer. to collect bacteria from solid agar medium, please don't mix bacterial suspension with liquid Agar medium, but just spread the cell suspension on solidified agar surfaface without breaking the agar surface with a suitable glass device, so that you can collect the cells after growth by pouring sterile saline water and then gently shaking the agar plate on your lab table or gently scrubing the bacterial colonies with an inoculation loop or any other smooth device.
Good luck.
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