Lidstrom: Tecan Plate Reader
From OpenWetWare
Back to Protocols
About the Tecan Infinite f500 plate reader
- This plate reader can do fluorescence tests.
- Product literature, machine specs.
Fluorescence-based promoter tests
- If you have promoters in front of a fluorescent protein and want to test the promoter strength. The answer you get will depend (nonlinearly!) on:
- The OD of the sample matters
- The gain you chose matters
- Should you subtract out fluorescence of the no-promoter control or divide the test value by the control?
- Evidence of both can be found in the literature
- You may have negative values for some samples.
- Is there a way to blank it??
- Evidence that the gain and the OD of the sample matter:
- You should definitely:
- Compare the test to a control at the same OD.
- This will require doing calculations for dilutions. You cannot just assume that 5uL of each type of cells + x uL water/buffer will give you the same OD.
- Things you should probably do:
- put cells in a non-fluorescent buffer.
- LB is known to contain fluorescent molecules.
- put cells in a non-fluorescent buffer.