Download Glycerol storage of bacterial cultures

Glycerol storage of bacterial cultures
1. To 0.85 ml [3 ml] of bacterial culture, add 0.15 ml [0.53 ml] of sterile glycerol
(sterilized by autoclave, final glycerol concentration : 15%, v/v).
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.
4. Freeze the culture in dry ice or in liquid nitrogen, and then transfer the tube to –70 C
for long-term storage.
* To recover the bacteria :
1) Scrape the frozen surface of the culture with a sterile inoculating loop.
2) Immediately streak the bacteria that adhere to the loop onto the surface of an LB
agar plate containing the appropriate antibiotics.
3) Return the frozen culture to storage at –70 C.
4) Incubate the plate overnight at 37 C.
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Dr. Lee’s Lab