Cell Preparation

 

Materials:

Methods:

1. Obtain a cell density of
2. Chill cells on ice
3. Washing procedure:
   1. Pellet 1 L of culture at 15 min
   2. Wash 1: Resuspend 1 volume sterile cold . Pellet 15 min
   3. Wash 2: Resuspend 0.5 volume sterile cold . Pellet 15 min
   4. Wash 3: Resuspend 0.5 volume sterile cold . Pellet 15 min
   5. Wash 4: Resuspend 0.02 volume sterile cold . Pellet 15 min
4. Final dilution: resuspend 0.002 - 0.003 volume sterile cold 10% Glycerol.
5. Aliquots may be frozen at , thaw on ice 5-10 min