Preparation of Bacterial Freezer Stocks

Materials

  • LB broth
  • Cryogenic tubes
  • 50% glycerol (autoclaved)

Protocol

  1. Pipette 5-10 mL LB (with appropriate antibiotic) into 15mL tube. For instructions on preparing LB broth, refer to Zamanian Lab Recipes.

  2. Inoculate a single colony from the streak or transformation plate.

  3. Set tube to shake in 37°C incubator overnight.

  4. After ~16 hours, take culture tubes out of the incubator and make freezer stocks as directed below.

  5. Label cryogenic tubes appropriately with a printed label.

  6. Add 500 μl of sterile 50% glycerol and an equal volume of culture to each cryo tube.

  7. Place tubes in correct -80°C and liquid N2 Bacterial Stock.

  8. Inventory freezer stocks in Quartzy.

See Lab Manual for complete tube labeling, tube count, and Quartzy inventory instructions.