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Team:ESBS-Strasbourg/7 August 2008
From 2008.igem.org
Contents |
DryLab
Modeling
WetLab
Paul
-recuperation of the overnight digest
-Gel migration and purification of the inserts c1/lacI/lexA/tetR/EYFP/adh1P/adh1t
-Dephosphorylation of the vectors containing respectively NLS/linker/Kozak sequences
precipitation phenol-chloroform and with alcool, resuspended in TE
vector +linker = 20µL (50ng/µL)
vector +NLS = 40µL (100ng/µL)
vector +Kozak = 500µL (50ng/µL)
-ligation for BB assembly:
ratio 1:3
1)c1/linker
2)lacI/linker
3)lexA/linker
4)tetR/linker
5)NLS/adh1-ter
6)EYFP/linker
7)adh1-prom/Kozak
ratio 1:8
respectively same construct than previously
8/9/10/11/12/13/14
plates with only the vectors dP, no insert: linker/Kozak/NLS
-Transformation in LB plates + Amp, overnight
