I've been trying to recover some bacterial cells by streaking them on agar plates, without success.
The stock has been kept in good conditions. But, there is a white pellet at the end of the glycerol tube, which I'm not actually sure whether it's the bacterial cells or not.
Do bacterial cells form pellets at the end of the glycerol stock tube?
Is there any way to recover them from the glycerol stock?
Dear Paul you should try to grow them in suitable broth medium first, and then transfer on agar plate. You can then identify if that is your organism or not. Though I never experience such condition with my glycerol stock, but may be possible in a longer duration.
Which technique of revival have you used? Did you freeze the bacterial samples with glycerol (-80C)?
Try to white heat microbiological loop and then quickly immerse in the frozen tube and make a streak on the plate.
Mujo Hasanović Yes, the bacterial samples were mixed with glycerol and stored at -80C.
I always took the sample from the top of the glycerol stock with my loop. Did you mean I need to take it from deeper parts of the stock?
Paul Weber Yes you need to immerse your loop deep into the tube. I worked for me and it will for you I hope so.
Yasemin Küçükata Thanks for your answer. Doesn't vortexing lyse the bacterial cells?
Yasemin Küçükata could you tell me how to repeat broth step more than once?
Should I just take some of the broth and add it to a new broth, and then incubate it again?
How much old broth should I use?
How long should I incubate the new broth?
Revival could require repeated subculture. Start by scrapping growth on the old slant into fresh tryptic soya broth and incubate in a shaking incubator till shows turbitity. Check for pure culture the revival depends on the type of microorganism and stage of preservation. Best of luck
Wafaa Eltayeb thanks for your comment. I actually use LB to culture my bacteria. But, does takeing some of the liquid culture and mixing it with a fresh liquid medium lead to a higher growth rate?
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