Thursday, June 22, 2006

Induction!

I innoculated two Novagen Overnight Express Auto Induction 125 ml cultures. As explained by Novagen,
The Overnight Express™ Autoinduction Systems enable regulated protein expression in E. coli without monitoring the culture or adding inducer during cell growth. These unique culture media are based on technology by F. William Studier at Brookhaven National Laboratory. ... Cell mass and target protein yield are often increased severalfold as compared with conventional protocols using induction with IPTG.


Hopefully my cultures are clean; I didn't use my clean air hood. I also skipped a staging step -- Novagen suggests starting with 30 mls and working up to 125 mls, but I'm trying to finish this by Saturday morning before I leave for a month for Africa.

Novagen recommends 16 hours of incubation at 37 before harvest. The flasks are on my orbital-hybrid microtitter shaker table at only 30 degrees; I plan to incubate overnight at this lower temperature. Novagen's product bulletin also says that the lower temperature may have other benefits. Cultures will have been incubating for 16 hours by 8:30 tomorrow; I'll harvest around noontime.

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