Team:Wash U/Protocol
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:Note: Restriction sites E=EcoR1-HF; X=Xba1; S=Spe1; P=Pst1; M=Mixed Site | :Note: Restriction sites E=EcoR1-HF; X=Xba1; S=Spe1; P=Pst1; M=Mixed Site | ||
:To view the full BioBrick Manual procedures, please click [http://partsregistry.org/Help:Transformation_Protocol here]. | :To view the full BioBrick Manual procedures, please click [http://partsregistry.org/Help:Transformation_Protocol here]. | ||
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'''Procedures''' | '''Procedures''' | ||
:List of Procedures | :List of Procedures | ||
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:3. Ligate the products. The result will be the upstream part connected to the downstream part in the destination plasmid. This new composite part can then be used to transform competent cells.<br> | :3. Ligate the products. The result will be the upstream part connected to the downstream part in the destination plasmid. This new composite part can then be used to transform competent cells.<br> | ||
:To view the full Biobrick Assembly manual, please click [http://ginkgobioworks.com/support/ here]. | :To view the full Biobrick Assembly manual, please click [http://ginkgobioworks.com/support/ here]. | ||
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:6. Transfer the tubes to an incubator set at 80C for another 20 minutes. This step will deactivate the restriction enzymes.<br> | :6. Transfer the tubes to an incubator set at 80C for another 20 minutes. This step will deactivate the restriction enzymes.<br> | ||
:7. Digestion is now finished and products should be stored at -20C.<br> | :7. Digestion is now finished and products should be stored at -20C.<br> | ||
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:5. Let the mix stand for 10 minutes at room temperature before incubating at 80C for 20 minutes (deactivates enzymes).<br> | :5. Let the mix stand for 10 minutes at room temperature before incubating at 80C for 20 minutes (deactivates enzymes).<br> | ||
:6. Store the products at -20C until they are needed for a transformation.<br> | :6. Store the products at -20C until they are needed for a transformation.<br> | ||
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'''Procedures''' | '''Procedures''' | ||
:text | :text | ||
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== '''Recipes''' == | == '''Recipes''' == | ||
<font size="2"> | <font size="2"> | ||
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'''Lysogeny Broth (LB) Media''' | '''Lysogeny Broth (LB) Media''' | ||
*10g Tryptone | *10g Tryptone |
Revision as of 16:14, 6 July 2009