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	::1. Growth O/N for 15h in LB (5ml) of:		::1. Growth O/N for 15h in LB (5ml) of:
-	- trasformed E.Coli with appropriate antibiotic;	+	-trasformed E.Coli with appropriate antibiotic;
-	- E. Coli.	+	-E. Coli.
	::2. All misure has to be done in M9 at OD=1.		::2. All misure has to be done in M9 at OD=1.

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Revision as of 10:17, 4 October 2007

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.