Janet B. Matsen:Growing Cultures For Crude Extract: Difference between revisions
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(New page: Return to Janet's Protocols == growing culture == # streak fresh plates from freezer stock # inoculate overnight cultures for subsequent inoculation # i...) |
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== storing cells == | == storing cells == | ||
# pour into 2 50 mL flasks | # pour into 2 50 mL flasks | ||
# centrifuge for | # centrifuge for 20 min at 4oC & 5000 rpm (add 10 min if needed) | ||
# flash freeze with liquid N2 | # flash freeze with liquid N2 | ||
# store @ -20oC | # store @ -20oC | ||
Revision as of 11:45, 29 May 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
- 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
- pour into 2 50 mL flasks
- centrifuge for 20 min at 4oC & 5000 rpm (add 10 min if needed)
- flash freeze with liquid N2
- store @ -20oC
