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Team:KULeuven/17 September 2008
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We started the analysis of the degradation of GFP-LVA for the thirth time, but this time with excellent results. Yesterday, we made liquid cultures of GFP, GFP-LVA and pUC. We re-inoculated them into fresh LB medium this morning and allowed the cells to grow for about 4 hours - this to make sure that the cells were in the log-phase when we start the measurements. Then we transferred the cells into a minimal ABT medium. This medium does not contain sugars or any other source of carbon. This way the cells will stop producing GFPs and so we can measure degradation. From the moment that we transferred the cells into this ABT medium, we started to measure the fluorescence of the cells with a FACS apparatus. We measured the fluorescence every 15 to 20 minutes for about 4 hours. The results were promising. The GFP with LVA tag showed a strong decline in fluorescence (as we expected) and the normal GFP was still very fluorescent after 4 hours. | We started the analysis of the degradation of GFP-LVA for the thirth time, but this time with excellent results. Yesterday, we made liquid cultures of GFP, GFP-LVA and pUC. We re-inoculated them into fresh LB medium this morning and allowed the cells to grow for about 4 hours - this to make sure that the cells were in the log-phase when we start the measurements. Then we transferred the cells into a minimal ABT medium. This medium does not contain sugars or any other source of carbon. This way the cells will stop producing GFPs and so we can measure degradation. From the moment that we transferred the cells into this ABT medium, we started to measure the fluorescence of the cells with a FACS apparatus. We measured the fluorescence every 15 to 20 minutes for about 4 hours. The results were promising. The GFP with LVA tag showed a strong decline in fluorescence (as we expected) and the normal GFP was still very fluorescent after 4 hours. | ||
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=== Dry Lab === | === Dry Lab === | ||
Started to make the lay-out of the notebook a little bit better. | Started to make the lay-out of the notebook a little bit better. | ||
Latest revision as of 01:48, 30 October 2008
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Lab Work
Wet Lab
We made a miniprep of the electroporations that succeeded: [http://partsregistry.org/Part:BBa_J23109 J23109]+[http://partsregistry.org/Part:BBa_J23078 J23078], [http://partsregistry.org/Part:BBa_K145253 K145253]+[http://partsregistry.org/Part:BBa_K145254 K145254], [http://partsregistry.org/Part:BBa_K145014 K145014] and [http://partsregistry.org/Part:BBa_K145201 K145201].
We started the analysis of the degradation of GFP-LVA for the thirth time, but this time with excellent results. Yesterday, we made liquid cultures of GFP, GFP-LVA and pUC. We re-inoculated them into fresh LB medium this morning and allowed the cells to grow for about 4 hours - this to make sure that the cells were in the log-phase when we start the measurements. Then we transferred the cells into a minimal ABT medium. This medium does not contain sugars or any other source of carbon. This way the cells will stop producing GFPs and so we can measure degradation. From the moment that we transferred the cells into this ABT medium, we started to measure the fluorescence of the cells with a FACS apparatus. We measured the fluorescence every 15 to 20 minutes for about 4 hours. The results were promising. The GFP with LVA tag showed a strong decline in fluorescence (as we expected) and the normal GFP was still very fluorescent after 4 hours.
Dry Lab
Started to make the lay-out of the notebook a little bit better.
