Making DMSO stocks of African Tryps (BSF and PCF)

Nathan Hutchings and Paul Grandgenett 8-9-00

Add ½ culture volume of fresh SM to your culture.

Spin culture for 10 minutes @ 2000 x g.

Resuspend pellet in 1/10 volume of SM minus HIFCS

Add equal volume of Freezing solution**

Mix gently by pipetting

Alliquot 500 ul aliquots into sterile 2 ml screw cap tubes.

Store tubes @ -80 C.

**Freezing Solution (SM 20% HIFCS + 15% DMSO)

Make 20 ml of SM 20% HIFCS. (16 ml SM + 4 ml of HIFCS)

Take 17 ml of the SM + 20%HIFCS and add 3 ml of tissue culture grade DMSO.

Mix thoroughly prior to adding to cells.