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Team:DTU-Denmark-2/Project/achievements
From 2011.igem.org
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| + | <b>Background</b><br> | ||
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| + | Even though we first started the iGEM project in the end of June, we have managed to design a fully standardized cloning system called Plug'n Play with DNA. We think that iGEM should be about fast and easy assembling of BioBricks, which led us to use features from the USER fusion standard assembly to model our <a hret="https://2011.igem.org/Team:DTU-Denmark-2/Project/overview"> Plug'Play with DNA </a>assembly system. Our system consist of: | ||
| + | <li> 49 BioBricks and 21 plasmids- All ready to use!. | ||
| + | <li> Back-up plasmide - To ensure mutation free amplification. | ||
| + | <li> Guide on customization - All procedures only require 1 round of PCR and assembly | ||
| + | <br><br> | ||
| + | We have characterized the two fungal promoters, PalcA and DMKP-P6, and three mammalian promoters, SV40, PGK, CMV. <br> | ||
| + | The <a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Characterisation">characterization</a> of the fungal promoters was qualitative evaluated with an X-gal analysis, where the expression of ß-galactosidase results in blue colonies. Furthermore, quantitative ß-galactosidase and Bradford assay was performed to measure the protein production and ß-galactosidase activity.<br> | ||
| + | To demonstrate Plug' Play with DNA also was applicable in <a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/mammalian">mammalian cell</a> lines, the transfected cells capability to express the fluorescence molecules was investigated with confocal microscopy.<br> | ||
| + | Both the characterization and fluorescence verified that the system worked as well in mammalian cells than fungi. | ||
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Revision as of 18:03, 16 September 2011
Background
Even though we first started the iGEM project in the end of June, we have managed to design a fully standardized cloning system called Plug'n Play with DNA. We think that iGEM should be about fast and easy assembling of BioBricks, which led us to use features from the USER fusion standard assembly to model our Plug'Play with DNA assembly system. Our system consist of:
We have characterized the two fungal promoters, PalcA and DMKP-P6, and three mammalian promoters, SV40, PGK, CMV.
The characterization of the fungal promoters was qualitative evaluated with an X-gal analysis, where the expression of ß-galactosidase results in blue colonies. Furthermore, quantitative ß-galactosidase and Bradford assay was performed to measure the protein production and ß-galactosidase activity.
To demonstrate Plug' Play with DNA also was applicable in mammalian cell lines, the transfected cells capability to express the fluorescence molecules was investigated with confocal microscopy.
Both the characterization and fluorescence verified that the system worked as well in mammalian cells than fungi.
