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

We are in Week 9  of the iGEM project

Project Calendar
name=IGEM:IMPERIAL/2007/Calendar date = 2007/08/01 view=threemonths format=%name/%year-%month-%day weekstart=7

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