CHP - last updated: January 29, 1999
This strain will not suppress the ambers so the protein or structure of interest will accumulate.
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.
Phage are so small in comparison to the cell this should give at least 1 phage/cell while others may get 10 phage/cell
Plate for bacterial survival Dilute 10-4 0.1 for colonies
Grow for 3 hours at 300C (or other more permissible temperature)
Before harvesting, take a few mls
If the cells are small and do not lysis there is a problem. Consider aborting the prep.
Spin culture - in centrifuge bottles - at 5K rpm for 5 minutes 4K/20 mins RC2C +
resuspend pellet in buffer B with 40mM octoglucoside.
Can begin freeze/thaw cycles of purification procedure