Stock Culture Preparation (Glycerol Stock) for Bacteria-A complete Procedure (ISO 11133:2014)
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- เผยแพร่เมื่อ 31 ต.ค. 2020
- Glycerol stock is very common and widely used procedure for the storage of microorganisms. This video represents a detailed procedure of Stock Culture for bacteria.
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Am very glad! The video is well explained. Thank you so much!
Thank you for such a detailed video....
Thank you for this Amazing video. Please make a video about bacterial biochemical tests.
Already uploaded one video of biochemical confirmation of bacterial. Please check our video list
👏👏👏👏👏👏Great
Hi! Thank you for this detailed video. My question is did you have problems with autoclaving glycerol? Some authors warn that glycerol will turn into acrolein if autoclaved, although it's uncertain whether it happens during short programs.
@Olga Radulovic, I appriciate your research. But in ISO method, it is said to use glycerol after autoclaving. You never know the glycerol, you want to use without autoclaving, is sterile and thus without sterilization your reference culture may become contaminated. So we must use sterile glycerin solution always.
We do practice autoclaving in our lab facing no major problem. If you get commercial sterile glycerine from a authentic & reliable source then you can use it without autoclaving.
Nice video , even we do culture revival in lab and before final preservation centrifuge 2-3 times so that culture gets in form of pellet and discard the broth liquid and finally add glycerol and store the pellet and store at -20 deg . I think centrifugation is mandatory . Any suggestions ??
You are correct
thank you for this amazing video❤ can you please tell me how to revive those cultures ?
Purchase from Sigma Aldrich
Excellent video the only negative point of this video is you used the eppendorf tubes
while in case of Glycerol stocks scientists don't recommend use of eppendorf as they can open at low unknowingly so use the stock tubes
and one more thing you can store the glycerol stock at -80° C for years
Great point! Stock tubes were not available to us at that time.
Anyways, thanks for the info.
Though thta's true, sometimes one don't have Cryovials at the lab. We instead have opted for using parafilm as sealant and let me tell you it worked pretty well, at least for 2 yrs.
Only when cryovials are not available
@@santiagoalvarez8993 This information will surely help me and others in future. Thanks ;)
Hai, thank you, wonderful video ,i need the reference for this ,, kindly guide me...i have to document alos
Excellent work
Thank you
@@MicroChemsExperiments can you please tell me what is final concentration of glycerol (in percentage) in final stock? and how to find it? as glycerol conc. was 20% initially.
Thanks in advance.
@@soulmeditationandhealing1074 Glycerol in final stock is 14%. It involves multiple calculation of percentage. Find it by yourself
Hi can we make liquid culture media from streak culture of ecoli?
Hi! Thank you for the video. The question is can I store bacteria in glycerol stock at -20 celsius degrees? Are there any negative effects?
Yes. You can store with no negative effect
Very useful video. I appreciate your great effort . But the Background music should not be so loud .
Thank you Ma'am.
We will make background music lower from next video
Can we use lactose broth instead TSB?
hi, i just want to ask, what is the ratio of your glycerol stock culture?
If we keep them at - 20 degrees, won't they die or contaminated? Caz preserve culture normally at - 70 noh?
can we add equal volume of overnight culture and glycerol?
No. Do as described in this video.
Thank you, is this preserving method valid for all types of bacteria?
Yes, it is
Or for filamentous fungi...?
Yes also used for fungi....@@shumaylakhan1220
Can we use nutrient broth with 10 percent glycerol and store at -20 fridge for a year
You can
What percent of glycerol can use in bile esculin agar? Plz reply
Please take a look at media catalogue
Thanks for this nice explanation.
I have questions if you didn’t mind
I will bay TSB with 15% glycerol broth from company it will come sterilised , which better to put the ATCC strain stick directly to this broth then after incubation I can transfer 2 ml from this broth to cryovials then stored at -80 degrees Celsius?
Or I must culture stick on agar then take from colonies to TSB with 15% glycerol broth then incubate them then transfer 2 ml from broth to cryovials?
How I can use TSB with 15% glycerol to preserve ATCC strain for years ?
Is it right to transfer TSB with 15% glycerol after put ATCC strain in it for cryovials??
Ay music too loud
Aaawww sorry for that...
We reduced music volume for other videos.