Team:Freiburg/Protocols/Cryo Stocks

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E. coli glycerol stocks

adapted from: Addgene-Glycerol stocks

material: E. coli o/n culture, 50% Glycerol, cryovial, liquid nitrogen
15 Min

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1. add 1000 μL of the overnight culture to 300μL of 50% glycerol in a labbeled cryovial and mix gently (by pipetting up and down)
2. freeze at -80°C with liquid nitrogen
3. store at -80°C
4. to recover bacteria from your glycerol stock, open the tube and use a sterile loop, toothpick or pipette tip to scrape some of the frozen bacteria off of the top. Do not let the glycerol stock unthaw!
5. Streak the bacteria onto an LB agar plate/liquid LB-medium
6. let it grow o/n

protocols, cloning