|
[Day 1]: Mummies Guide to Making Broth! Objectives: To describe the steps to prepare a bioreactor To prepare the media for seed culture and scale fermentation To prepare the seed culture for scale up fermentation Procedures: 1. Media for Seed-Culture and Scale-up Fermentation Before we actually begin the fermentation process, we have to prepare the media for which the cells will be grown in, the same way you cannot swim in a swimming pool if there is no water inside. Also, unlike us, cells “eat” the media, so we have to make sure that the media has the necessary ingredients for their “all their nutritional needs” *Baby milk powder advertisement voice*.   1. Prepare 2.0 litre of Luria-Bertani Medium (LB),  Indeed, these cells that we grow are very much like  4. Add ampicillin to a final concentration of 100mg/ml  2. Bioreactor Preparation Steps1. Calibrate the pH electrode using electrode using standard buffer solution (pH 7 and pH 4 or 9 depending on the culture). 2. Install the pH probe, pO2 probe, foam and level probe into the top plate. 3. Connect the addition agent lines for acid, base and antifoam. Check the levels in the storage bottles. 4. Install other accessories such as exhaust condensers, air inlet and exhaust filters and manual sampler unit. Check that the water jacket is filled with water. 5. Prepare for sterilization: i) Disconnect all cables except the temperature probe, which is autoclavable. ii) Clamp all silicone tubings except for exhaust filter and female STT coupling of sampling unit. iii) Cover all filters and sockets with aluminium foil to protect from condensing moisture. iv) Autoclave with steam at 121°C for 20 minutes. 6. Polarise the pO2 electrode for at least 6 hours. Calibrate it by aerating with nitrogen.
7. Connect the addition lines to peristaltic pumps. Switch to “Auto” or “Manual” control appropriately.  We have spoken.
Comments:
发表评论
|
 |
