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Team:Calgary/Notebook/Protocols/MiniExpression
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<li>Make 2mL overnight cultures of your protein-producing bacteria</li> | <li>Make 2mL overnight cultures of your protein-producing bacteria</li> | ||
<li>Subculture in 10mL of LB broth</li> | <li>Subculture in 10mL of LB broth</li> | ||
| - | + | <li>When OD600 is between 0.3 and 0.5, induce the culture with IPTG</li> | |
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<li>To determine optimum conditions for expression, use different time point of induction (e.g. 4h, 8h and 12h) and different concentration of IPTG (e.g., 0.1mM, 0.4mM and 0.8mM)</li> | <li>To determine optimum conditions for expression, use different time point of induction (e.g. 4h, 8h and 12h) and different concentration of IPTG (e.g., 0.1mM, 0.4mM and 0.8mM)</li> | ||
</ol> | </ol> | ||
Latest revision as of 09:03, 27 September 2013
Mini Expression
Mini Expression
We used this protocol to define the optimum conditions for expression of our constructs.
Reagents and Materials
- Overnight cultures of protein-producing bacteria
- LB broth
- IPTG (Isopropyl-Beta-D-Thiogalactoside)
Protocol
- Make 2mL overnight cultures of your protein-producing bacteria
- Subculture in 10mL of LB broth
- When OD600 is between 0.3 and 0.5, induce the culture with IPTG
- To determine optimum conditions for expression, use different time point of induction (e.g. 4h, 8h and 12h) and different concentration of IPTG (e.g., 0.1mM, 0.4mM and 0.8mM)
