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From 2008.igem.org

Contents 1 Glycerol Stocks 1.1 Materials 1.2 Method 1.3 Notes

Glycerol Stocks

Materials

• 40% glycerol solution
• Cryogenic vials

Method

1. Add 1 ml of 40% glycerol in H2O to a cryogenic vial.
2. Add 1 ml sample from the culture of bacteria to be stored.
3. Gently vortex the cryogenic vial to ensure the culture and glycerol is well-mixed.
   • Alternatively, pipet to mix.
4. Use a tough spot to put the name of the strain or some useful identifier on the top of the vial.
5. On the side of the vial list all relevant information - part, vector, strain, date, researcher, etc.
6. Store in a freezer box in a -80C freezer. Remember to record where the vial is stored for fast retrieval later.

Notes

While it is possible to make a long term stock from cells in stationary phase, ideally your culture should be in logarithmic growth phase. Prof. Schonbaum's version

1. Grow a 3 ml culture overnight
2. Add 200 µl 100% glycerol to 1 ml cells in a cryotube (final concentration should be 15-20% glycerol different concentrations from different protocols
3. Mix by vortexing and let sit for a little bit. Then put the tubes in the -80°C freezer.

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