a. Open the vial according to the enclosed instructions.
b. Using a single tube of broth (5 to 6 mL), withdraw approximately 0.5 to 1.0 mL with a Pasteur or
1.0 mL pipette. Rehydrate the entire pellet.
c. Aseptically transfer this aliquot back into the tube. Mix well.
d. Make serial dilutions by transferring 0.5 mL from the original tube to a tube containing 4.5 mL.
Repeat process by transferring 0.5 mL from the second to a third tube, etc. Dilutions are
important, not only for titration purposes, but also to keep culture in varying stages of growth.
Many strains will die out rapidly once acid or alkaline conditions are reached. It is recommended
to prepare several dilutions from the initial tube as the cryoprotectant used in the freeze