Imperial/Wet Lab/Lab Notebook/2007-08-15


15 August 2007

Preparation of Cell Extract

  1. Innoculated 45 ml of fresh overnight culture in 1 litre 2xYT medium
  2. Incubated at 37°C until O.D. 600 = 0.600
  3. Added 5 ml of 1 mM IPTG
  4. Incubated at 37°C
    • O.D. 600 was 1.543 at 6pm
  5. Culture left at 37° in the incubator overnight
  6. Incubated new 100 ml culture overnight

Midiprep of Biobricks

  1. Midipreped 9 Biobricks

  • 2. BBa_I13422 [ptet-GFP]
  • 5. BBa_R0040 [ptet]
  • 7. BBa_F2620 [plux]
  • 8. BBa_F1610 [luxI]
  • 11. BBa_E7104 [pT7]
  • 13. BBa_B0015 [stop]
  • 14. BBa_R0051 [pcI]
  • 17. BBa_I13507 [RFP]
  • 18. BBa_T9002 [plux-GFP]

Protocols can be found at Midiprep in the general protocols page

Pilot Preparation of Vesicles

  • Results:

The vesicles prepared the day before were looked at once again under a light microscope, with phase contrast. This time, vesicles were observed, with size range 2-43μm.

IC07 image077.jpg IC07 image078.jpg IC07 image079.jpg
Photos taken with a portable camera through the microscope eye-piece. Setup: 100x oil immersion objective, with phase contrast.

  • Preparations: The desiccation and suspension stages of the vesicle preparation protocol were carried out once again.
  1. One 100ml beaker was prepared with 125μl of DOPC in 50ml of mineral oil
  2. The suspension was sonicated for 30mins and left overnight in a 27°C incubator
  • It was later found that the desiccator had no vacuum