Bologna University/Fluorescence issue
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< Bologna University
Revision as of 10:14, 4 October 2007 by Silvia.tamarri2 (Talk | contribs)
- 1Growth O/N for 15h in LB (5ml) of:
- trasformed E.Coli with appropriate antibiotic; - E. Coli.
2. All misure has to be done in M9 at OD=1. 3.The day after in the morning OD measure. 4. To obtain OD=1:
-centrifugate 4ml of bacteria culture at 4400rpm for 3min at 25°C; -eliminate the supernatant; -risuspende the pellet with 6ml in M9 medium with glucose with appropriate antibiotic (1000x).
5. OD measure: adjust OD to 1 through further diluition or growth. 6. Adjust PMT offset with untrasformed E.Coli. 7. Fluorescence test of the culture before IPTG induction. 8. IPTG induction:
-centrifugate bacteria culture at 4400rpm for 3min at 25°C; -eliminate the supernatant; -risuspende the pellet in M9 medium and 2mM IPTG with appropiate antibiotic (1000X);
9. Incubate at 37°C.
10. Fluorescence test after10min, 20min, 30min, 1h, 2h.