Making Glycerol Stocks Protocol

For single use stocks: follow steps 2 - 5 but instead of step 6, pipet 50 – 100 ul of cells to sterile labeled microfuge tubes. Note that if you don’t have many cells, you can reduce the volume of MHB and glycerol (keep the same ratio, with a final concentration of glycerol of 15%. E.g. 800 uL MHB and 200 uL 75% glycerol).

Make 3 cryovials for each strain (permanent stocks), label!

Prepare 2.4mL of MHB in a 50mL conical (adjust if you are also making single use stocks)

Take at least half of a thickly spread plate and add cells to the MHB tube

Resuspend until there are no clumps in the MHB

Add 600ul of 75% glycerol to the 2.4mL mix by pipetting

Aliquot 1mL per cryovial, freeze at -80