Bologna University/Fluorescence issue

From 2007.igem.org

(Difference between revisions)
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::::-centrifugate bacteria culture at 4400rpm for 3min at 25°C;
::::-centrifugate bacteria culture at 4400rpm for 3min at 25°C;
::::-eliminate the supernatant;
::::-eliminate the supernatant;
-
::::-risuspende the pellet in M9 medium and 2mM IPTG with appropiate antibiotic (1000X);
+
::::-risuspende the pellet in M9 medium and 2mM IPTG with appropiate antibiotic(1000X);
::9.  Incubate at 37°C.
::9.  Incubate at 37°C.
::10. Fluorescence test after10min, 20min, 30min, 1h, 2h.
::10. Fluorescence test after10min, 20min, 30min, 1h, 2h.

Revision as of 10:20, 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.