IGEM:IMPERIAL/2007/Calendar/2007-9-5

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