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	::10. Test fluorescence after 10min, 20min, 30min, 1h, 2h.		::10. Test fluorescence after 10min, 20min, 30min, 1h, 2h.
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Latest revision as of 09:43, 24 October 2007

1. Growth O/N for 15h in LB (5ml) of:

-transformed E. Coli with appropriate antibiotic;

-E. Coli.

2. All measures have to be done in M9 with OD=1.2.

3. The day after in the morning measure OD.

4. To obtain OD=1.2:

-centrifugate 4ml of bacteria culture at 4400rpm for 3min at 25°C;

-discard the supernatant;

-resuspende cell pellet in 6ml of M9 medium with glucose and appropriate antibiotic (1000x).

5. Measure OD: adjust OD to 1.2 through further diluition or growth.

6. Adjust PMT offset with untransformed E. Coli.

7. Test culture fluorescence before IPTG induction.

8. IPTG induction:

-centrifugate bacteria culture at 4400rpm for 3min at 25°C;

-discard the supernatant;

-resuspende cell pellet in M9 medium and 2mM IPTG with appropriate antibiotic (1000X);

9. Incubate at 37°C.

10. Test fluorescence after 10min, 20min, 30min, 1h, 2h.

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