From 2007.igem.org

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Revision as of 10:14, 4 October 2007

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.