Making Protein in the Phage system

CHP - last updated: January 29, 1999


Step 1


Step 2 

Before infection, take a sample of the culture examine under the microscope. The cells should look like sausages and should be swimming.

T=0 plate for colonies at 10-6 dilution 0.1 ml.

Step 3


Step 4
Plate for bacterial survival Dilute 10-4 0.1 for colonies


Step 5
Grow for 3 hours at 300C (or other more permissible temperature)


Step 6
Before harvesting, take a few mls

If the cells are small and do not lysis there is a problem. Consider aborting the prep.


Step 7
Spin culture - in centrifuge bottles - at 5K rpm for 5 minutes 4K/20 mins RC2C +


Step 8
resuspend pellet in buffer B with 40mM octoglucoside.


Step 9
Can begin freeze/thaw cycles of purification procedure