Bologna University/Fluorescence issue

From 2007.igem.org

1. Growth 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.