Now the Battle with I0500 Begins...
- Kanamycin plates with pUC and I0500 TOP10 cells did not give any colonies
- Incubation time with SOC for 2 hours is too short!
- Plate out again with TOP10 incubated overnight at 37°C
- TOP10 transformed with J63006 plated out on ampicillin plates (neat and 1/10 dil)
- pUC control with 1/10 dil also plated out on ampicillin plates
- Incubated at 37°C overnight
Anaerobic Condition and Iron Assay
- Require nitrogen gas and paraffin oil
- Received paraffin oil from the Pathology Dept, return excess after use
- Nitrogen gas available in the Plant Dept
- Dye for iron ions to be ordered from Sigma
- Fur- mutant single colony plated on Kanamycin plate and inoculated into 75mL M9 medium prepared on 23/7/08
- Mutants in M9 incubated at 37°C overnight