This protocol may be usedµ to make a glycerol stock of a bacterial strain.

Keywords: bacteria glycerol

Category
Protocol
Record History
Added on May 3, 2010 at 9:41 AM by Lindner, Jill
Modified on July 1, 2010 at 2:13 PM by Lindner, Jill
Shared with (contributions)
VCSCBi: Training / Protocols Master

Reagents, supplies, and equipment:

  • Luria-Bertani broth (LB)
  • 75% glycerol
  • 1.5 ml Eppendorf tubes
  • Nunc tubes

Procedure:

  1. Inoculate 5 ml LB with a single colony from an overnight plate or with 5 µl of mini-prep DNA bacterial culture and grow overnight.
  2. Add 800 µl of the fresh overnight culture + 200 µl 75% glycerol in a clean 1.5 ml Eppendorf tube; mix thoroughly with pipette.
  3. Transfer to Nunc tube, and store at -70°C.

Written by Kathy Shelton