<a href="https://2011.igem.org/Team:DTU-Denmark-2/Project/achievements#Fulfilment of medal requirements" class="h1"> Fulfilment of medal requirements</a><br><br>
The team came together during a course in mammalian cell biology in June, and that's when the idea of iGEM popped into our heads. Eventhough we started the iGEM project in late June, we have managed to design a fully standardized cloning system called Plug'n Play with DNA. We thought that time should be spent on making cool iGEM systems, instead of struggeling with unwanted restriction sites and limited assembly systems. This led us to the USER fusion standard assembly as a model our <a hret="https://2011.igem.org/Team:DTU-Denmark-2/Project/overview"> Plug'Play with DNA </a>assembly system.</p><br>
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The team came together during a course in mammalian cell biology in June, and that was when the idea of iGEM popped into our heads. Even though we started the iGEM project late June, we have managed to design a fully standardized cloning system called Plug'n Play with DNA. We thought that time should be spent on making cool iGEM projects, instead of struggling with unwanted restriction sites and cumbersome assembly systems. This led us to standardize USER fusion by designing the<a hret="https://2011.igem.org/Team:DTU-Denmark-2/Project/overview"> Plug'n'Play with DNA </a>assembly system.</p><br>
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<li> 49 BioBricks and 21 plasmids- All ready to use!.</li>
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<li> 49 ready-to-use BioBricks.</li>
<li> 21 backup plasmids.</li>
<li> 21 backup plasmids.</li>
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<li> A quick 'n' easy guide on how to custumize new BioBricks or introducing specific mutations</li>
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<li> A quick 'n' easy guide on how to customize the standard</li>
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To ensure that the BioBricks and devices have been assembled correctly we have tested the functionality of them in fungi and in mammalian cells.
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To ensure that the BioBricks and devices have been assembled correctly we have tested the functionality of them in <i>Aspergillus nidulans</i> and in mammalian cells.
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We have characterized the two fungal promoters, PalcA and DMKP-P6, and proved the function of three mammalian promoters, SV40, PGK, and CMV.
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We have characterized the two fungal promoters, P<i>alcA</i> and DMKP-P6, and demonstrated function of the cytomegalovirus (CMV) promoter in mammalian cells.
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The <a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Characterisation">characterization</a> of the fungal promoters was successfully evaluated with an X-gal analysis, where the expression of ß-galactosidase results in blue colonies. Furthermore, quantitative ß-galactosidase and Bradford assays were performed in order to measure protein production and ß-galactosidase activity. Both promoters proved to be of medium strength.
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The <a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Characterisation">characterization</a> of the fungal promoters was successfully performed with an X-gal analysis, where the expression of ß-galactosidase results in blue colonies. Furthermore, quantitative ß-galactosidase and Bradford assays were performed in order to measure protein production and ß-galactosidase activity. Both promoters proved to be of medium strength.
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For demonstration of the Plug' Play with DNA system in <a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/mammalian">mammalian cell</a> lines, U-2 OS was transfected with plasmids containing genes for fluorescence molecules. The transfected cells capability to express the fluorescence molecules was investigated with confocal microscopy, which resulted in amazing <a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/mammalian">pictures</a> of glowing peroxisomes and colored U-2 SO cells.
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For demonstration of the Plug' Play with DNA system in <a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/mammalian">mammalian cell</a> lines, U-2 OS was transfected with plasmids containing genes encoding fluorescent proteins. The transfected cells capability to express the fluorescent proteins was investigated with confocal microscopy, which resulted in amazing <a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/mammalian">pictures</a> of fluorescent peroxisomes and coloured U-2 SO cells.
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All the submitted BioBricks have been sequenced, and the data have been examinated in order to verify that no mutations or incorrect insertions had occurred. The analysis of the sequencing data showed that the Plug 'n' Play parts had been correctly assembled. The sequencing and the characterization both show that the system works as expected in fungi and mammalian cells without unwanted insertion and mutations.
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Some of the submitted BioBricks have been successfully sequenced, and the data has been examined in order to verify that no mutations or incorrect insertions had occurred. The analysis of the sequencing data showed that all the sequenced Plug 'n' Play BioBricks had been correctly assembled without mutations. The sequencing and the characterization both showed that the system works as expected in <i>A. nidulans</i> and in the mammalian cell line U-2 OS.
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<a name="Fulfilment of medal requirements"></a><h1><b>Fulfilment of medal requirements</b></h1>
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<b><h3>Medal accomplishment</h3></b>
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<b><big>Bronze</big></b><br>
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<b>1. Team Registration</b><br>
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All team members are registrated <br><br>
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<b>2. Complete judging form </b><br>
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Judgingform will be completed as soon as possible.<br><br>
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<b>3. Create and share a description of the teams project using iGEM wiki and the Team's parts.</b><br>
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We have made a description of our project (see The project), and submitted parts can be found under Results.<br><br>
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<b>4. Plan to present a poster and talk at the iGEM Jamboree.</b><br>
We have made a description of our project (see The project), and submitted parts can be found under Results.<br><br>
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<input type="checkbox" checked disabled="true"><b> Present a poster and a talk at the iGEM Jamboree</b><br>
We plan to make an excellent presentation and a magnificent poster that will be transported on first class from Denmark to Amsterdam right before the Jamboree. <br><br>
We plan to make an excellent presentation and a magnificent poster that will be transported on first class from Denmark to Amsterdam right before the Jamboree. <br><br>
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<input type="checkbox" checked disabled="true"> <b>At least one new submitted and well-characterized standard BioBrick Part or Device. A new application of and outstanding documentation (quantitative data showing the Part’s/ Device’s function) of a previously existing BioBrick part in the “Experience” section of that BioBrick’s Registry entry also counts.</b><br>
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We have submitted 49 new <a hret="https://2011.igem.org/Team:DTU-Denmark-2/results/submitted_biobricks">BioBricks</a>.
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<b>5. Enter information detailing at least one new standard BioBrick Part or Device in the Registry of Standard Biological Parts</b><br>
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We have submitted 49 new <a hret="https://2011.igem.org/Team:DTU-Denmark-2/results/submitted_biobricks">BioBricks</a>.
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<b>Silver</b><br> <br>
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<input type="checkbox" checked disabled="true"><b>Demonstrate that at least one new BioBrick Part or Device of your own design and construction works as expected</b><br>
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We have demonstrated the function of the device, <a hret=!http://partsregistry.org/Part:BBa_K678002"> BBa_K678002</a>, that has been transfected into the mammalian cell line, U-2 OS. Furthermore, characterization of fungal promoter parts have been executed. These results can be viewed at our homepage or in the BioBrick registry for <a href="http://partsregistry.org/Part:BBa_K678000">BBa_K678000</a> and <a href="http://partsregistry.org/Part:BBa_K678001">BBa_K678001</a><br><br>
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<input type="checkbox" checked disabled="true"><b>Characterize the operation of at least one new BioBrick Part or Device and enter this information in the “Main Page” section of that Part’s/Device’s Registry entry.</b><br>
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<b>Gold</b> <br>
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<b><big>Silver</big></b><br>
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<input type="checkbox" checked disabled="true"><b>Improve the function of an existing BioBrick Part or Device (created by another team or your own institution in a previous year) and enter this information in the Registry (in the “Experience” section of that BioBrick’s Registry entry), and don't forget to create a new registry page for the improved part.</b><br><br>
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<b>1+2. Demonstrate that at least one new BioBrick Part or Device of your own design and construction works as expected; characterize the operation of your new part/device. Enter this information on the "Main Page" section of the registry </b><br>
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We have demonstrated the function of the device, <a hret=!http://partsregistry.org/Part:BBa_K678002"> BBa_K678002</a>, that has been transfected into the mammalian cell line, U-2 OS. Furthermore, characterization of fungal promoter parts have been executed. These results can be viewed at our homepage or in the BioBrick registry for <a href="http://partsregistry.org/Part:BBa_K678000">BBa_K678000</a> and <a href="http://partsregistry.org/Part:BBa_K678001">BBa_K678001</a>
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<b><big>Gold</big></b><br>
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<b>1:Improve an existing BioBrick Part or Device and enter this information back on the Experience Page of the Registry.</b><br>
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We used the CMV promoter, <a href="http://partsregistry.org/Part:BBa_J52034"> BBa_J52034 </a>, as the regulator for our fluorescence reporters, and it worked well. The promotor and our experience is described in <a href="http://partsregistry.org/Part:BBa_J52034:Experience"> Experience</a>. <br><br>
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<b>2: Help another iGEM team by, for example, characterizing a part, debugging a construct, or modeling or simulating their system. </b><br>
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We have helped the iGEM team: Copenhagen with the design and assembly of some of their <a href="https://2011.igem.org/Team:Copenhagen/Project/Data">BioBricks</a>. <br><br>
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<input type="checkbox" checked disabled="true"><b>Help another iGEM team by, for example, characterizing a part, debugging a construct, or modeling or simulating their system.</b><br>We have helped the iGEM team Copenhagen with the design and assembly of some of their <a href="https://2011.igem.org/Team:Copenhagen/Project/Data">BioBricks</a>.<br><br>
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<input type="checkbox" disabled="true"><b>Outline and detail a new approach to an issue of Human Practice in synthetic biology as it relates to your project, such as safety, security, ethics, or ownership, sharing, and innovation.</b><br><br>
The team came together during a course in mammalian cell biology in June, and that was when the idea of iGEM popped into our heads. Even though we started the iGEM project late June, we have managed to design a fully standardized cloning system called Plug'n Play with DNA. We thought that time should be spent on making cool iGEM projects, instead of struggling with unwanted restriction sites and cumbersome assembly systems. This led us to standardize USER fusion by designing the Plug'n'Play with DNA assembly system.
Our Plug 'n' Play with DNA kit consist of:
49 ready-to-use BioBricks.
21 backup plasmids.
A quick 'n' easy guide on how to customize the standard
To ensure that the BioBricks and devices have been assembled correctly we have tested the functionality of them in Aspergillus nidulans and in mammalian cells.
We have characterized the two fungal promoters, PalcA and DMKP-P6, and demonstrated function of the cytomegalovirus (CMV) promoter in mammalian cells.
The characterization of the fungal promoters was successfully performed with an X-gal analysis, where the expression of ß-galactosidase results in blue colonies. Furthermore, quantitative ß-galactosidase and Bradford assays were performed in order to measure protein production and ß-galactosidase activity. Both promoters proved to be of medium strength.
For demonstration of the Plug' Play with DNA system in mammalian cell lines, U-2 OS was transfected with plasmids containing genes encoding fluorescent proteins. The transfected cells capability to express the fluorescent proteins was investigated with confocal microscopy, which resulted in amazing pictures of fluorescent peroxisomes and coloured U-2 SO cells.
Some of the submitted BioBricks have been successfully sequenced, and the data has been examined in order to verify that no mutations or incorrect insertions had occurred. The analysis of the sequencing data showed that all the sequenced Plug 'n' Play BioBricks had been correctly assembled without mutations. The sequencing and the characterization both showed that the system works as expected in A. nidulans and in the mammalian cell line U-2 OS.
Fulfilment of medal requirements
Bronze
Team registration
Complete Judging form
Team Wiki
We have made a description of our project (see The project), and submitted parts can be found under Results.
Present a poster and a talk at the iGEM Jamboree
We plan to make an excellent presentation and a magnificent poster that will be transported on first class from Denmark to Amsterdam right before the Jamboree.
At least one new submitted and well-characterized standard BioBrick Part or Device. A new application of and outstanding documentation (quantitative data showing the Part’s/ Device’s function) of a previously existing BioBrick part in the “Experience” section of that BioBrick’s Registry entry also counts.
We have submitted 49 new BioBricks.
Silver
Demonstrate that at least one new BioBrick Part or Device of your own design and construction works as expected
We have demonstrated the function of the device, BBa_K678002, that has been transfected into the mammalian cell line, U-2 OS. Furthermore, characterization of fungal promoter parts have been executed. These results can be viewed at our homepage or in the BioBrick registry for BBa_K678000 and BBa_K678001
Characterize the operation of at least one new BioBrick Part or Device and enter this information in the “Main Page” section of that Part’s/Device’s Registry entry.
Gold
Improve the function of an existing BioBrick Part or Device (created by another team or your own institution in a previous year) and enter this information in the Registry (in the “Experience” section of that BioBrick’s Registry entry), and don't forget to create a new registry page for the improved part.
Help another iGEM team by, for example, characterizing a part, debugging a construct, or modeling or simulating their system. We have helped the iGEM team Copenhagen with the design and assembly of some of their BioBricks.
Outline and detail a new approach to an issue of Human Practice in synthetic biology as it relates to your project, such as safety, security, ethics, or ownership, sharing, and innovation.
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