Team:Cambridge/LRMeetingNotes
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Meeting with Laura Rowe Notes
- Brightness measurements are integrated over time, so when comparing them make sure they are integrated over a long time (otherwise you are only measuring how quickly luminescence occurs, not how bright).
- Different colours:
- Could use fluorophores instead of fluorescent proteins but need to ensure they attach to protein (could be a project in itself). Requires resonance energy transfer.
- Using mutant luciferases probably easier to implement.
- Expression in E. coli: inclusion bodies could be a problem when trying to over-express genes. To test: break open cells and centrifuge twice (at a higher speed second time around), if pellets are formed these are probably inclusion bodies. Could then dissolve these with solvent and test for bioluminescence from proteins within inclusion bodies.
- Relative light units are used because different camera properties, distances etc. all mean that photons/sec are relative. Can use a source for calibration - tritium standards are used (e.g. MGM instruments).