Team:Cambridge/Notebook/Week2

Test

Alex Orda at [http://www.anachem.co.uk/ Anachem] for the free supply of EarthSaver pipette tips, freezer boxes and the very snazzy IsoFreeze racks.

Monday

Did further research into the enzymes required for luciferase recovery.

• Presented proposals to advisors
  • Modelled RNA to create aptazyme based system using trial of CLC RNA Workbench
  • Made presentation for Newcastle
  • Set up and tested low light camera

Tuesday

• 06.50 - Train to Newcastle for UK iGEM get-togther

Wednesday

Thursday

• 11am - Health and Safety talk by Barbara
• Working lunch at the Waffle Company. Summary of discussion:
  • To do:
    • Ordering
    • Getting in touch with MIT (turning off lights)
    • T-Shirts
    • Project Plan (Gantt Chart etc)
  • To do long-term:
    • Modelling
    • Firefly Bioluminescence
    • Bacterial Bioluminescence
    • Human Practices
    • Quiescence
  • Individual roles:
    • Outline of experiments, protocols - Anja, Ben, Peter (especially controls)
    • Modelling - Paul, Bill, Emily
    • COSHH forms - Bill
    • LRE biobrick - Theo
    • Researching Bacterial Lux operon - Will, Hannah

Friday

Meeting with Laura Rowe in the Biotechnology Dept. at 3pm:

• 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).

Paper from Duncan about mutant bacterial strain with very bright luminescence to be investigated further.

Saturday

Sunday