Team:Northwestern/Protocols/Glycerol

From 2012.igem.org

Glycerol Stocks

Materials

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

Procedure

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