Imperial/Wet Lab/Lab Notebook/2007-08-15
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| - | Protocols can be found at [ | + | Protocols can be found at [http://www.openwetware.org/wiki/IGEM:IMPERIAL/2007/Notebook/General_Protocols#Midiprep Midiprep] in the general protocols page |
==Pilot Preparation of Vesicles== | ==Pilot Preparation of Vesicles== | ||
Revision as of 15:42, 25 October 2007
Contents |
15 August 2007
Preparation of Cell Extract
- Innoculated 45 ml of fresh overnight culture in 1 litre 2xYT medium
- Incubated at 37°C until O.D. 600 = 0.600
- Added 5 ml of 1 mM IPTG
- Incubated at 37°C
- O.D. 600 was 1.543 at 6pm
- Culture left at 37° in the incubator overnight
- Incubated new 100 ml culture overnight
Midiprep of Biobricks
- 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 [http://www.openwetware.org/wiki/IGEM:IMPERIAL/2007/Notebook/General_Protocols#Midiprep 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.
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| 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.
- One 100ml beaker was prepared with 125μl of DOPC in 50ml of mineral oil
- The suspension was sonicated for 30mins and left overnight in a 27°C incubator
- It was later found that the desiccator had no vacuum
