Difference between revisions of "Team:UCLA/Notebook/Honeybee Silk/12 August 2015"
(→Multiple Mini Expressions: Optimizing protein yield) |
(→Multiple Mini Expressions: Optimizing protein yield) |
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*We will test these different conditions using the following protocol | *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) | #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. | + | #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. | ||
Revision as of 22:15, 20 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.