Bacterial glycerol stocks

Virtualab Protocols (Chang Bioscience)
1. Grow 10 ml overnight liquid culture with antibiotics selection.

2. Sterilize 30% (v/v) glycerol by autoclave.

3. Add 0.5 ml bacterial culture to 0.5 ml 30% (v/v) glycerol in 1.5 ml screw-capped vial or eppendorf tubes.

4. Invert tubes to mix cells and glycerol, then store at -80 C.

5. Sterile DMSO can be used instead of glycerol.

6. To revive stored cells, scraping off splinters of solid ice with a sterile toothpick or inoculation loop and streaking onto a LB plate. Avoid complete thawing of the stock.