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  • Making Glycerol Stocks

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    htrakr
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    To prepare -80°C glycerol stocks for long-term storage of bacterial cultures grown in liquid media, add 0.5 ml of sterile 60% glycerol to 1.5 ml of bacterial culture, vortex to mix, and freeze in liquid nitrogen before storing at -80°C. Bacteria can be recovered by scraping the frozen surface with a sterile loop and streaking onto agar plates or into liquid media.

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    To prepare -80°C glycerol stocks for long-term storage of bacterial cultures grown in liquid media, add 0.5 ml of sterile 60% glycerol to 1.5 ml of bacterial culture, vortex to mix, and freeze in liquid nitrogen before storing at -80°C. Bacteria can be recovered by scraping the frozen surface with a sterile loop and streaking onto agar plates or into liquid media.

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    14 views1 page

    Making Glycerol Stocks

    Uploaded by

    htrakr
    To prepare -80°C glycerol stocks for long-term storage of bacterial cultures grown in liquid media, add 0.5 ml of sterile 60% glycerol to 1.5 ml of bacterial culture, vortex to mix, and freeze in liquid nitrogen before storing at -80°C. Bacteria can be recovered by scraping the frozen surface with a sterile loop and streaking onto agar plates or into liquid media.

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    Preparation of -80C Glycerol stocks Storage of Bacterial Cultures Growing in Liquid Media 1. To 1.5 ml of bacterial culture, add 0.

    5 ml of sterile 60% glycerol (sterilized by autoclaving for 20 minutes at 15 psi on liquid cycle). 2. Vortex the culture to ensure that the glycerol is evenly dispersed. 3. Transfer the culture to a labeled storage tube equipped with a screw cap and an air tight gasket. 4. Freeze the culture in ethanol-dry ice or in liquid nitrogen and then transfer the tube to -80C for long term storage. 5. To recover the bacteria, scrape the frozen surface of the culture with a sterile inoculating loop, and then immediately streak the bacteria that adhere to the needle onto the surface of an LB agar plate/or in a 5ml LB tube containing the appropriate antibiotic. Stocks die quickly at room temp so if you need to take it out of the freezer, be sure to keep on ice.

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