Kay Finn
Last revision: 5/13/2003
1) Glycerol solution:
65% glycerol (vol/vol) (52 ml glycerol)
0.1 M MgSO4 (8ml of 1M stock)
0.025 M Tris-Cl pH 8 (2ml of 1M Tris-Cl)
pH with NaOH to pH 8
...add sterile H2O to 80 ml total volume
2) Autoclave this solution and have sterile cryo vials (screw cap, thick walled)
ready (labeled, etc.).
3) Have cells in mid log phase (best) or fresh stationary phase.
4) Add 10 ml of glycerol solution to15 ml of mid-log cell culture, mix well
and fill cryo tubes with 1.5 ml aliquots.
5) Store in Baker/Clark –80ºC freezer in equipment room. Update Freezer
Box Database, and make sure box/area is dated and labeled.