Day 2 - Ready... Get Set... Media and Seed Culture Preparation...

Objective

• To prepare seed culture for scale-up fermentation.

B. Bioreactor Preparation Steps

1. pH probe, pO2 probe, foam and level probe was installed into the top plate.
   
2. The agent lines for acid, base and antifoam was connected and the storage bottles was checked. Other equipment was also installed such as exhaust condensers, air inlet and exhaust filters.
   
3. All the filters and sockets were covered with aluminum foil to protect from condensing moisture. Cables were all disconnected except the temperature probe, which is autoclaved. All silicone tubings except for exhaust filter and female STT coupling of sampling unit.
   
4. pO2 electrode was polarized for at least 6 hours and the pO2 probe was calibrated by aerating with oxygen.

C. Preparation of Seed Culture

1. pGLO transformed E.coli was retrieved from -800C.
   
2. The E.coli was streak on a LB/Amp/Ara plate and incubate for 24 hours.
   
3. Several colonies of pGLO transformed E.coli was taken and transfer into a flask containing 100ml LB medium with ampicillin.
   
4. The flask was put in shaking incubator and was incubate at 320C for 24 hours and which is was used to inoculate the fermenter for scale-up fermentation.

Streaking the E.Coli. JunJie getting into action...

Making sure they are alive and kicking...... well not literally kicking, ...but you get the idea...!

Sadha stealing the limelight from JunJie.......