Team:UCLA/Notebook/Honeybee Silk/12 August 2015
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
- Grow up a starter culture using glycerol stock #1 of our silk in BL21 to an OD of around 1. (4 to 5 hours)
- Add 50 ul of starter culture to 5 ml of LB starting around 5 pm along with 5 ul of Kan.
- Add in the varying amounts of IPTG and use the two different temperatures.