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From 2010.igem.org

Freezing of bacterial stocks

Materials:

- bacterial culture

- LB medium

- 80% glycerol

- centrifuge

Protocol:

1. Take 5 mL bacterial cells from the Erlenmeyer of a freshly grown culture and spin in a 15 mL tube for 10 minutes at 2,000 rpm (Eppendorf centrifuge).

2. Decant the supernatant without disturbing the pellet.

3. Pipet on the pellet 0.5 mL 1% LB medium and 0.5 mL 80% glycerol and mix by vortexing and save in -80 °C freezer.