Lidstrom:Autoinduction Media: Difference between revisions
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==Recipe: == | ==Recipe: == | ||
* [[image:150101_Autoinduction_recipe_from_internet.png|upright= | * [[image:150101_Autoinduction_recipe_from_internet.png|upright=2.0|thumb|center|Autoinduction Media recipe from Studier 2005.]] | ||
* [[image:150101_Autoinduction_media_antibiotic_specifications.png|upright=2.0|thumb|center|Autoinduction Media antibiotic specificiations.]] | |||
== Why Autoinduction Media? == | == Why Autoinduction Media? == | ||
Revision as of 10:44, 1 January 2015
Return: Protocols
Recipe:
Autoinduction Media recipe from Studier 2005.
Autoinduction Media antibiotic specificiations.
Why Autoinduction Media?
Instead of inducing a T7 promoter with IPTG, autoinduction media can be used. Glucose, glycerol, and fructose are added to a buffered yeast broth to make autoinduction media. As the E. coli cultures grow, they consume the glucose first, then as they are forced to use the fructose, the T7 promoter is turned on and your T7-driven gene is expressed.
The advantages are:
- you don't have to monitor OD of the cultures
- you can go straight from -80oC to protein purification in 1 day instead of two
- great for library screening where cultures have different growth rates
References
- Original Paper: Studier FW (2005), Protein Production by Auto-Induction in High-Density Shaking Cultures. Protein Expr. Purif. 41(1): 207–234. <-- we make media ZYM-5052.
- Practical summary slides: Imperial college .ppt slides: http://www3.imperial.ac.uk/pls/portallive/docs/1/15699698.PPT
Comparing autoinduction media to TB induced with IPTG
Janet did an experiment in 12/2014 comparing cell pellets of His-tagged ADH grown in autoinduction media to the same culture grown in TB and induced with IPTG as part of this experiment. The autoinduced culture had comparable amounts of protein as seen by SDS-PAGE, and yielded more protein upon Co-NTA purification.
