Cryopreservation
Dilute 100 % Glycerol to 50 % Glycerol .
In a sterile 50ml Falcon tube pour 25ml 100 % glycerol .
Add 25ml autoclaved MilliQ and mix thoroughly .
Pour into sterile reservoir .
In sterile 500ml deep 96 well plate add 240μl of 50 % glycerol to each well using an 8 channel pipet .
Take pore film off of 2ml deep 96 well plate .
Using a 12 channel 200μl pipet , pipet up and down from bottom of wells 3 times , then transfer 160μl from each well to the plate with glycerol .
Pipet up and down in glycerol 5 times .
Repeat step 4 for all rows / wells .
Seal 500ml deep 96 well plate ( glycerol plate ) with foil .
Place in shaker for 30 minutes .
Re - seal 2ml deep 96 well plate with pore film and place in refrigerator .
After 30 minutes on shaker , place 500ml deep plate in - 80ºC freezer
