[Glycerol stocks] plasmids

Published on Author Anne Petzold

Instructions according to Stockinger lab manual: http://www.oardc.ohio-state.edu/stockingerlab/Protocols/GlycerolStocks.pdf

Make a glycerol stock of plasmid

  • pick a single colony from the transformation plate and grow an o/n culture in appropriate medium, e.g. LB amp
  • put 0.5 ml of 80 % sterile glycerol in sterile screw cap microcentrifuge tube
  • add 0.5 ml of o/n culture
  • vortex
  • freeze @ -80 DEG
  • check with a digestion of a miniprep whether stock construct is correct

Take sample from glycerol stock

  • take a sterile blue plastic scraper and scrape off a portion from the top of the stock
  • plate the portion on an agar plate and grow o/n
  • DON’T LET STOCK THAW!

 

80 % glycerol stock:

  • 80 % glycerol
  • 20 % ddH2O