Stock Culture Preparation (Glycerol Stock) for Bacteria-A complete Procedure (ISO 11133:2014)

[@fightfanatic9102]

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

[@MicroChemsExperiments]

Great point! Stock tubes were not available to us at that time. Anyways, thanks for the info.

[@santiagoalvarez8993]

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

[@fightfanatic9102]

@@santiagoalvarez8993 This information will surely help me and others in future. Thanks ;)

[@segunabisoye]

Beautiful presentation, please I would like to ask if is a must to use diluents prepared instead of just distilled water especially working with nutrient agar and the compatible salt/butter advice. Kindly guide me.

[@dgi3947]

Am very glad! The video is well explained. Thank you so much!

[@muhammadzubair929]

Thank you for such a detailed video....

[@soniapradhan1095]

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 ??

[@MicroChemsExperiments]

You are correct

[@shyanaali1134]

Thank you for this Amazing video. Please make a video about bacterial biochemical tests.

[@MicroChemsExperiments]

Already uploaded one video of biochemical confirmation of bacterial. Please check our video list

[@EelasMusic]

If we keep them at - 20 degrees, won't they die or contaminated? Caz preserve culture normally at - 70 noh?

[@ugurcangul4792]

Hi nice video.I have question.if i need spesific concentration like 30 cfu for qc what steps should i take from here

[@dr.urmibhattacharjee6919]

Very useful video. I appreciate your great effort . But the Background music should not be so loud .

[@MicroChemsExperiments]

Thank you Ma'am. We will make background music lower from next video

[@maram1maram]

You made a big mistake the final concentration of glycerol in the glycerol stock became 14% u have to use 60% first and just 600uL from it to get a final concentration of 30% because the final concentration of glycerol have to be from 20% to 40% maximum

[@sumalathamanju7191]

Hai, thank you, wonderful video ,i need the reference for this ,, kindly guide me...i have to document alos

[@olgaradulovic7684]

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.

[@MicroChemsExperiments]

@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.

[@Ney194]

thank you for this amazing video❤ can you please tell me how to revive those cultures ?

[@MicroChemsExperiments]

Purchase from Sigma Aldrich

[@PhúcHuỳnhDuy-c8r]

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?

[@MicroChemsExperiments]

Yes. You can store with no negative effect

[@sadiaurmila3745]

pls avoid the background music for better learning

[@Sjjeien]

Hi can we make liquid culture media from streak culture of ecoli?

[@nurayozsoy690]

Can we use lactose broth instead TSB?

[@asem7611]

Thank you, is this preserving method valid for all types of bacteria?

[@MicroChemsExperiments]

Yes, it is

[@shumaylakhan1220]

Or for filamentous fungi...?

[@MoazzamRajpoot-sw4ys]

Yes also used for fungi....​@@shumaylakhan1220

[@soulmeditationandhealing1074]

Excellent work

[@MicroChemsExperiments]

Thank you

[@soulmeditationandhealing1074]

@@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.

[@MicroChemsExperiments]

@@soulmeditationandhealing1074 Glycerol in final stock is 14%. It involves multiple calculation of percentage. Find it by yourself

[@akramasgari]

👏👏👏👏👏👏Great

[@WallaceDaveHajjyPLim]

hi, i just want to ask, what is the ratio of your glycerol stock culture?

[@aftabkhan1189]

3:7 (Culture : 20% Glycerol)

[@jeenathapa6146]

can we add equal volume of overnight culture and glycerol?

[@MicroChemsExperiments]

No. Do as described in this video.

[@ANIRUDH774]

Can we use nutrient broth with 10 percent glycerol and store at -20 fridge for a year

[@MicroChemsExperiments]

You can

[@nahidayashmin6084]

What percent of glycerol can use in bile esculin agar? Plz reply

[@MicroChemsExperiments]

Please take a look at media catalogue

[@batoolalardah8123]

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??

[@kabelomatlalambono4189]

Ay music too loud

[@MicroChemsExperiments]

Aaawww sorry for that... We reduced music volume for other videos.