Day 2
Experiment 3: Inoculation, Fermentation and Monitoring
Objectives
1. To carry out scale-up fermentation process for GFP protein
2. To monitor cell growth and product formation through manual sampling and computer data logging
Procedure for setting up the fermentation process (Part I):
1. The ampicillin and arabinose stock solution had been prepared before hand and filtered and sterilized.
2. The medium was cooled to below 50°C. Ampicillin was added until it reached a final concentration of 100μg/ml, while arabinose was added until it reached a final concentration of 0.2%.
3. The following parameters were set up:
pH - 7.5
Stirring speed - minimum 10%, maxiumum 90%, control to AUTO
pO2 - 20%, control to AUTO, stir to "CASC" & "AIRFLOW" to "CASC"
Airflow - minimum 25%, maximum 100%
4. The fermentor was inoculated with 100ml of seed culture (5% of fermentation medium volume).
5. The fermentation was then continued for another 24 hours.
Back to Homepage
Day 1, Experiment 1: Familiarization of bioreactor and equipmentsDay 1, Experiment 2: Equipment and media preparationDay 3: Sampling and monitoring of cell growthDay 4: Harvesting the cellsDay 5: Isolation and purification of product