Janet B. Matsen:Growing Cultures For Crude Extract: Difference between revisions
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== growing culture == | == growing culture == | ||
Revision as of 16:20, 6 June 2012
Return to Janet's Protocols
growing culture
- streak fresh plates from freezer stock
- inoculate overnight cultures for subsequent inoculation
- inoculate into 100 mL of M9 media (or 50 mL of TB)
- grow to mid-exponential at 37oC, shaking at 225 rpm
- add inducers (aTc & IPTG) and shake at 18oC & 200 rpm for 36 hours!
storing cells
- split each between 2 50 mL flasks
- centrifuge for 20 min at 4oC & 5000 rpm (add 10 min if needed)
- flash freeze with liquid N2
- store @ -80oC (she says you generally do) in a magic blue box
