These should be made with the rest of the overnight culture not used in the miniprep procedure.

• Overnight Cultures
• 60% Glycerol by volume (Autoclaved)
• 2mL microcentrifuge tubes

Create the Glycerol Stock

1. Create the Glycerol Stock
2. Add 1.5mL of the overnight stock into the microcentrifuge tube
3. Add .5mL of the sterile 60% glycerol stock to the microcentrifuge tube
4. Gently shake and mix to ensure even distribution of the glycerol
5. Label the tube, and put it into the -80C freezer

Recovering the Cells

1. Scrape the cells from the ice on the glycerol stock with a sterile loop
2. Plate the cells with the loop on a plate with antibiotics
3. Place the glycerol stock back in the -80C freezer
4. Incubate the plate at 37C overnight for individual colonies