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Fermentation (2)

Fermentation (2) - FERMENTATION Duration 2 weeks 4 lab...

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FERMENTATION Duration: 2 weeks, 4 lab sessions Week 1: Learn fermentor setup. Perform growth of expression strain. Week 2: Fermentation with protein expression, harvest and lysis. This section of the lab will allow you to directly work with the high cell density fermentation. Since cell growth in fermentation is very different than shake flask fermentation, we will spend the first week to understand fermentor assembly, study the growth pattern of the expression strain in the fermentor. In the second week, your group should be able to independently perform an entire fermentation run, including protein expression and cell harvest. There are two prelab assignments associated with the fermentation modules due during session 1 and session 3 respectively 1. Fermentor Basics 1. Draw a schematic diagram of the fermentor and label all components. 2. Derive an equation for calculating the doubling time and growth rate of E. coli . 2. Fermenting E. coli for Protein Expression 1. Plot the growth data obtained from previous fermentation and label different phases of growth. 2. Plot both OD measurements and DO levels. How do they correlate with each other? 3. Calculate the doubling time using the plot. Session 1: Fermentor assembly, sterilization and preculture preparation 1. Prepare 2 L LB medium (consult with TA), 300 mL NaOH [1M] and 300 mL HCl [1M]. Dissolve the LB on a stirrer. 2. Sterilize 400 mL of the LB medium, two test tubes and a shake flask with lids attached in the autoclave. Coordinate with the other groups for the use of the autoclave.
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