IGEM:IMPERIAL/2007/Notebook/2007-8-15

name=iGEM:IMPERIAL/2007/Notebook date=2007/09/15 view=threemonths format=%name/%year-%month-%day weekstart=7

Preparation of Cell Extract

 * 1) Innoculated 45 ml of fresh overnight culture in 1 litre 2xYT medium
 * 2) Incubated at 37&deg;C until O.D. 600 = 0.600
 * 3) Added 5 ml of 1 mM IPTG
 * 4) Incubated at 37&deg;C
 * 5) *O.D. 600 was 1.543 at 6pm
 * 6) Culture left at 37&deg; in the incubator overnight
 * 7) 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 2-43&mu;m.


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