Materials

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

Protocol

1. Pipette 5–10 mL LB with the appropriate antibiotic into a 15 mL tube. For instructions on preparing LB broth, see General Lab Recipes.
2. Inoculate a single colony from the streak or transformation plate.
3. Set the tube to shake in a 37°C incubator overnight.
4. After ~16 hours, take the 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 the correct -80°C and liquid N₂ bacterial stock boxes.
8. Inventory freezer stocks in Quartzy.

<aside> 🏷️

See the ‣ for complete tube labeling, tube count, and Quartzy inventory instructions.

</aside>