Team:UCLA/Notebook/Honeybee Silk/12 August 2015

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Multiple Mini Expressions: Optimizing protein yield

• Today we are testing multiple growth conditions of our bl21 cells for total cell and protein yield. Parameters we are optimizing are iptg concentration, and growth temperature.

IPTG concentration/Temp	37C	30C
0.1 mM
0.5 mM
1.0 mM
2.0 mM

• We will test these different conditions using the following protocol

1. Grow up a starter culture using glycerol stock #1 of our silk in BL21 to an OD of around 1. (4 to 5 hours)
2. Add 50 ul of starter culture to 5 ml of LB starting around 5 pm along with 5 ul of Kan.
3. Add in the varying amounts of IPTG and use the two different temperatures.