Purdue/11 July 2008

From 2008.igem.org

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Glycerol Stocks!

  • Today making glycerol stocks of SOS (J22106) and LacZ (I732017) parts from stabs sent by iGEM
  • We only have 100% glycerol (sterile), so we're just going to follow the same iGEM protocol but with 100% glycerol
  • Dr. Clase also suggested a somewhat different protocol for making stocks, so we're going to do both side-by-side and see which is better
  • Following iGEM protocol:
    • Make o/n culture in antibiotic (amp) LB broth --did this yesterday
    • Combine 1mL of overnight w/ 150 uL of sterile 100% glycerol solution in screw-top cryotube (make sure tube is labeled with part #, plasmid, and antibiotic resistance)
    • Vortex briefly
    • Incubate at room temp. for 1/2 hour and put in -80C freezer
  • Following Dr. Clase's protocol:
  • Mix 750uL 100% glycerol and 250uL stock colony in cryotube
  • Vortex
  • Put directly in -80C freezer

In other news, the bead plating did not help the other Lac transformations to work, although the pUC19 still grew quite well.

Edited by Janie Stine