Preparation of glycerol stocks

1. This protocol should only be used with strains that either do not sporulate (ie E. coli) or a strain that sporulates too poorly to collect spores from.
2. To sterile 2mL, screw-cap tubes filled with fresh, fairly high density, bacterial culture add 0.5 ml of sterile 60% glycerol.
3. Vortex to evenly disperse the glycerol.
4. Snap freeze in liquid nitrogen or a dry ice / ethanol bath (note that the ethanol will dissolve any label that you put on the tube).
5. Give to Susan along with a strain information form.
6. Glycerol stocks need to be stored at -80 C.

This protocol is taken directly from "Molecular Cloning - A Laboratory Manual", Third Edition. Sambrook and Russell, 2001, appendix A, section 8, page 5.