Team:ETH Zurich/Biology/Cloning
From 2011.igem.org
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(→PCR) |
(→Colonie PCR) |
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** annealing: 55 °C for 30 sec | ** annealing: 55 °C for 30 sec | ||
** extension: 68 °C for 90 sec | ** extension: 68 °C for 90 sec | ||
- | * final extension: | + | * final extension: 68 °C for 5 min |
- | * storage at | + | * storage at 4 °C |
== Whole plasmid PCR == | == Whole plasmid PCR == |
Revision as of 20:11, 19 September 2011
Material and methods |
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All protocols, plasmids and primers can be found here. |
ProtocolsDouble digest for cloning
Ligation
Transformation
PCRPCR mixture
adjust to 50 µl with ddH2O PCR procedure
Colonie PCRpick a colonie and resupsend in 10 µl ddH2O PCR mixture
adjust to 10 µl with ddH2O PCR procedure
Whole plasmid PCRPreparation of glycerol stocksAlcR testing
AntibioticsMediumsSOB
adjust to pH 7.5 by adding 1M NaOH SOC
LB medium
Total 1 l M9 minimal medium 10x M9
Total 100 ml autoclave seperatly
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Plasmid list |
Primer list |
Synthesized parts
- LacIM1
- AlcR (codon optimized)