Bologna University/Fluorescence issue

< Bologna University(Difference between revisions)

Latest revision as of 15:46, 4 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.

3. The day after in the morning measure OD.

4. To obtain OD=1:

-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 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.

@@ Line 11: / Line 11: @@
 ::::-discard the supernatant;
 ::::-resuspende cell pellet in 6ml of M9 medium with glucose and appropriate antibiotic (1000x).
-::5. measure OD: adjust OD to 1 through further diluition or growth.
+::5. Measure OD: adjust OD to 1 through further diluition or growth.
 ::6. Adjust PMT offset with untransformed E. Coli.
 ::7. Test culture fluorescence before IPTG induction.