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Team:ESBS-Strasbourg/7 August 2008
From 2008.igem.org
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'''Paul'''<br> | '''Paul'''<br> | ||
-recuperation of the overnight digest <br> | -recuperation of the overnight digest <br> | ||
| + | |||
-Gel migration and purification of the inserts c1/lacI/lexA/tetR/EYFP/adh1P/adh1t <br> | -Gel migration and purification of the inserts c1/lacI/lexA/tetR/EYFP/adh1P/adh1t <br> | ||
| + | |||
-Dephosphorylation of the vectors containing respectively NLS/linker/Kozak sequences <br> | -Dephosphorylation of the vectors containing respectively NLS/linker/Kozak sequences <br> | ||
precipitation phenol-chloroform and with alcool, resuspended in TE <br> | precipitation phenol-chloroform and with alcool, resuspended in TE <br> | ||
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vector +NLS = 40µL (100ng/µL) <br> | vector +NLS = 40µL (100ng/µL) <br> | ||
vector +Kozak = 500µL (50ng/µL) <br> | vector +Kozak = 500µL (50ng/µL) <br> | ||
| + | |||
-ligation for BB assembly: <br> | -ligation for BB assembly: <br> | ||
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plates with only the vectors dP, no insert: linker/Kozak/NLS <br> | plates with only the vectors dP, no insert: linker/Kozak/NLS <br> | ||
| + | |||
| + | |||
-Transformation in LB plates + Amp, overnight <br> | -Transformation in LB plates + Amp, overnight <br> | ||
Latest revision as of 09:54, 8 August 2008
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 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
