Gill:Making frozen permanents

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Revision as of 09:19, 10 September 2013 by Jacqueline K. Grimm (talk | contribs)


  • Cryo vials
  • Overnight bacterial culture
  • LB (50% glycerol)


Store plasmid-containing E. coli in 10% glycerol (200 μL LB (50% glycerol) added to 800 μL overnight culture in a cryo vial), and everything else in 15-20% glycerol. Vortex, put in a freezer box and store in the -80 degree freezer.


Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!

  • When making LB in 50% glycerol, measure out the glycerol in a graduated cylinder before adding water. To get the last of the glycerol out of the cylinder, add the required amount of water, cover the top with parafilm, and shake before pouring out.