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Team:Paris/3 August 2009
From 2009.igem.org
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Brain work
Meeting this morning for the lab work week. Discuss about the schedule algorithm for the iPhone application.
Lab work
DID dying for membrane fluorescence microscopy
- wash cell with MgSO4 then dying and incubation 30 min 25°C ->observation : 1/100 fluorescent cell and some vesicles ?!
- dying in medium, incubation 30min 37°C ->observation : no significant (1/100)
- dying in medium, incubation 30min 37°C, wash with MgSO4 ->observation : no significant (1/100)
->try another method : FM 4-64
Miniprep digestion by XbaI and PstI
Gel Migration
- [P1|P2|P3|P4|P5|Ladder 100bp|P7|P8]
- P1 = pSB2K3 = plasmid backbone = 4399bp (expected weight)
- P2 = BBa_J61002 = plasmid backbone = (will not be digest by this enzyme)
- P3 = BBa_J32015 = pelB = 66bp
- P4 = BBa_B0014 = Double terminater = 95bp
- P5 = BBa_I14032 = pLac = 37bp
- P7 = BBa_R0040 = tetR repressible promoter = 54bp (weak but present, cf other photos)
- P8 = BBa_I0500 = pBad/araC = 1210bp
Culture ON for Miniprep
- S7 DH5α(g3p)
To do list
| Matricule | TODO |
| Luc | SNARE Construction |
| Romain | Design oligo Fec A |
| Charlotte | design of oligos: FecR, FecA, SNARE?, sequencing oligo |
| Stoff | Dev schedule algorithm |
| Chris | modelling: Tol-Pal system, delay system, first equations in biocham ; use of mathlab. Think about the transduction system |
| Lisa | Oligo Tol/Pal / Oligo Sequencing / Tol/Pal Protocol verification / Keio / Theme A writting |
| Caroline | coloration of cells / protocols for the lab: control of the emission of vesicules.... |
| Souf | design of oligos: FecR, FecA, SNARE?, sequencing oligo |
| Vicard | planification of all the steps necessary for the implementation of the ClyA device in our strains / search for articles on fusion of vesicules |
| Pierre | modelling: Tol-Pal system, delay system, first equations in biocham ; use of mathlab. Think about the transduction system |
| Sylvain | minipreps. Check if the use of starch is possible ; problem : find a system to help the starch enter into the periplasm, use of LacY -> noise |
| Guillaume | miniprep digestion using Xba/Pst enzymes |
