Team:University of Chicago/Notebook/Glycerol stocks

From 2008.igem.org

Revision as of 18:50, 29 July 2008 by Norayucel (Talk | contribs)
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)

Contents

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.
Retrieved from "http://2008.igem.org/Team:University_of_Chicago/Notebook/Glycerol_stocks"