Team:ESBS-Strasbourg/7 August 2008

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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 named 8/9/10/11/12/13/14

plates with only the vectors dP, no insert: linker/Kozak/NLS

-Transformation in LB plates + Amp, overnight