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

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We are in Week 9 of the iGEM project

Project Calendar

July
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August
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September
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30


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
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