Can I make glycerol stocks of transformed Rosetta/BL21 cells?

[Muhammad Rahman]

Hi All,

Is it possible (or advisable) to make glycerols stocks of Rosetta/BL21(DE3) cells and store them? Also, when I have single colonies of transformed cells on agar plates, how long can I store them at 4C.

Your help/suggestion is appreciated,

Muhammad
University of Alberta

[dlcevo1]

It should be fine. I've kept BL21 and CL41 cells in glycerol stocks for storage in liquid N2 for over a year with no problems. I use a ratio of 1:1 glycerol to inoculum. As far as plates in the fridge, I've kept ioslated colonies viable for over 6 months, which is funny because my advisor told me 2-3 weeks max. The thicker the nutrient agar the better and make sure you wrap the plate with Parafilm to keep the moisture in.