Bologna University/Fluorescence Test
From 2007.igem.org
(Difference between revisions)
| Line 11: | Line 11: | ||
::::-discard the supernatant; | ::::-discard the supernatant; | ||
::::-resuspende cell pellet in 6ml of M9 medium with glucose and appropriate antibiotic (1000x). | ::::-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 1.2 through further diluition or growth. |
::6. Adjust PMT offset with untransformed E. Coli. | ::6. Adjust PMT offset with untransformed E. Coli. | ||
::7. Test culture fluorescence before IPTG induction. | ::7. Test culture fluorescence before IPTG induction. | ||
Revision as of 10:54, 19 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.
