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Team:Washington/Alkanes/Methods
From 2011.igem.org
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[[Image:Washington_2011_PetroBrick.png|220px|frameless|border="2"|link=http://partsregistry.org/wiki/index.php?title=Part:BBa_K590025|left]] | [[Image:Washington_2011_PetroBrick.png|220px|frameless|border="2"|link=http://partsregistry.org/wiki/index.php?title=Part:BBa_K590025|left]] | ||
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In order to produce alkanes, we need both acyl-ACP reductase (AAR) and aldehyde decarbonylase (ADC) to work together in the cell. In order to achieve this goal, we used standard cloning methods to construct a Biobrick that contained both AAR and ADC under a high constitutive promoter, each with it's own Elowitz standard RBS. This construct successfully synthesized our target product, and thus we have created a new modular alkane-producing platform: | In order to produce alkanes, we need both acyl-ACP reductase (AAR) and aldehyde decarbonylase (ADC) to work together in the cell. In order to achieve this goal, we used standard cloning methods to construct a Biobrick that contained both AAR and ADC under a high constitutive promoter, each with it's own Elowitz standard RBS. This construct successfully synthesized our target product, and thus we have created a new modular alkane-producing platform: | ||
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[[Image:Washington2011_PetroBrick_Construct.png|730px|frameless|border|left|link=http://partsregistry.org/wiki/index.php?title=Part:BBa_K590025]] | [[Image:Washington2011_PetroBrick_Construct.png|730px|frameless|border|left|link=http://partsregistry.org/wiki/index.php?title=Part:BBa_K590025]] | ||
Revision as of 22:09, 17 September 2011
Introducing the PetroBrick
In order to produce alkanes, we need both acyl-ACP reductase (AAR) and aldehyde decarbonylase (ADC) to work together in the cell. In order to achieve this goal, we used standard cloning methods to construct a Biobrick that contained both AAR and ADC under a high constitutive promoter, each with it's own Elowitz standard RBS. This construct successfully synthesized our target product, and thus we have created a new modular alkane-producing platform:
the PetroBrick.
Alkane Production & Extraction
After we had the complete gene assembly in our hands, the next step was to transform it into cells and start them growing for alkane production. We let them grow in 37 degree shaker for 48-72 hours, in sealed glass tubes. After the cells have gone through the alkane production process, the next step is to extract the alkanes out of the cell broth. We add acyl acetate directly into the glass test tube for cell growth. Then we vortex untill to everything is well mixed, to make sure all of the alkanes go directly into the ethyl acetate solvent. Next, we spin down the mixture by using a centrifuge at full speed to form three layers (cell pellet, media, and ethyl acetate supernatant). We use only the ethyl acetate layer to send for GCMS analysis.
Alkane Detection
Explain GCMS... again basically graphics from presentation with text
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