Template:Team:KULeuven/1 September 2009/BlueLightReceptor
From 2009.igem.org
- Experimental set up for irradiation of LigA ( + )
- Enting colonies from the motherplate:
- 4 agar plates with LigA = 2 for irradiation and 2 for control.
- 4 liquid cultures with LigA = 2 for irradiation and 2 for control.
- 4 agar plates with = 2 for irradiation and 2 for control.
- 4 liquid cultures with = 2 for irradiation and 2 for control.
- There are two groups, each with 8 cultures.
- Shift one: 2 agar plates and 2 liquid cultures with LigA and 2 agar plates and 2 liquid cultures with were put in 16°C for one hour. After an hour, half of them were irradiated with blue light for an hour while the other have was wrapped in aluminium foil as a control. Afterwards they are put in the 37°C incubator for some time
- Shift two: they were put in the 37°C for some hours so that the cultures can grow.
- After measurement of the first shift the following was concluded:
- Although the cells with had a small signal after FACS, the plates showed no GFP when put under the lamp.
- LigA that was not exposed to blue light had the same amount of signal as the construct that had been exposed. we assume that this is due to the exposure to white light when the cultures are exposed. Possibly, the promotor was then already activated since white light contains blue light frequencies. A new set up will be made over the weekend where all cultures will be made in a dark room.
- Dilutions of the shift one liquid cultures were made + 2 new cultures from the motherplate from ligA. These were put overnight under blue light together with the cultures of shift 1
- Following was ented in liquid culture:
- LigA from motherplate
- from motherplate
- DB3.1
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