IGEM:IMPERIAL/2007/Calendar/2007-9-5
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We are in Week 9 of the iGEM project
Project Calendar
<calendar> name=IGEM:IMPERIAL/2007/Calendar date = 2007/08/01 view=threemonths format=%name/%year-%month-%day weekstart=7 </calendar>
Debrief
Data Analysis
- Currently being carried out by the modelling team
- Some issues with staggered data and the pTet results
- Need to figure out why the results are so variant
- Can only carry on tests on pTet once reasons for the problems are figured out
- Degradation curves seem fine
- Still in the process of putting all the results on the wiki
- Some issues with the way the data is saved and caculations made in excel
- Modelling and experimental design team need to resolve these issues
Experiments
- Tested pLux construct with [AHL] of: 3nM, 5nM and 7nM
- Fluorescence seemed to be higher than in earlier experiments even though the concentrations of AHL were higher before
- May be as the DNA used earlier was mini-preped whereas that being used now has been midi-preped
- Need to look into it further
- Purification of LuxR - growing cells at the moment
- Will make the buffers tomorrow
- Might be done by Friday
Cloning
- pT7 has been cloned
- Quality check:
- In-vivo - works
- In-vitro - will be tested on Monday (DNA preparation - Friday)
Vesicles
- Home-made cell extract was put in to vesicles
- A lot of green fluorescence was seen but not sure if it was GFP being expressed
- Need working cell extract - exact amount needed to be calculated
- Can use the T7 cell extract and test the pT7 construct in the vessicles (as not enough of the commercial S30)
- Pore protein - found somewhere to buy from cheaply
- If use this one need to change the phospholipid currently being used - and the new phospholipid will need to be tested
- Otherwise need to keep looking for a new pore protein
GFP
- The GFP from regisrty E0040 is not wild type - actually a mutant: GFP-mut3b
- Can't get GFP-mut3b from GeneArt, thus can't get the right calibration and degradation curves