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Team:Penn/MaGellinToolbox
From 2013.igem.org
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| - | <header><h1><b><center>< | + | <header><h1><b><center>Toolbox</center></b></h1></header> |
| + | </br> | ||
| + | </br> | ||
| + | </br> | ||
| + | For a detailed, graphical explanation of the MaGellin work flow, please download the <a href="https://static.igem.org/mediawiki/2013/e/e5/Spec_Sheet.pdf">MaGellin Workflow Specifications Sheet</a>, which includes all of the steps in the MaGellin workflow. | ||
| + | </br> | ||
| + | </br> | ||
| + | </br> | ||
| + | <h4><b>Constructing a Toolbox</b></h4>Our goal was to create a toolbox for developing customized site-specific methylases. Like any good toolbox, it includes three components: the tool, the assay, and the automation. | ||
| + | <ol> | ||
| + | <li>The Tool: a fusion between a DNA binding domain and a methylase enzyme</li> | ||
| + | <li>The Assay: a digestion based assay, called MaGellin, to measure both methylase activity and sequence specificity</li> | ||
| + | <li>The Automation: a unique software package that predicts experimental outcomes and analyzes gel electrophoresis images to measure methylase functionality</li> | ||
| + | </ol> | ||
| + | </br> | ||
| + | </br> | ||
| + | <div style="margin-left:auto;margin-right:auto;text-align:center"><figure><img border="0" src="https://static.igem.org/mediawiki/2013/4/4b/Penn_Toolbox.png" alt="Toolbox" width="700" height="395"><figcaption><i>Toolbox with the three essential components.</i></figcaption></figure></div> | ||
| + | </br> | ||
| + | </br> | ||
| + | </br> | ||
| + | Our toolbox satisfies 6 requirements: | ||
| + | <ol> | ||
| + | <li>Robust – includes functional fusion proteins for comparison</li> | ||
| + | <li>Standardized – all inclusive in one-plasmid with simple multiple cloning sites, standard bisulfite sequencing primers, and a digestion based methylation assay built in</li> | ||
| + | <li>Easy to Use – we programmed a software package to predict experimental outcomes and automate gel electrophoresis analysis</li> | ||
| + | <li>Inexpensive – methylase activity and specificity can be screened by simply digesting and running a gel</li> | ||
| + | <li>Noiseless – the bacterial chassis has no background CpG methylation</li> | ||
| + | <li>Open Source – we<a href="https://2013.igem.org/Team:Penn/Biobricks"> BioBrick’ed</a> the full plasmid and the methylases</li> | ||
| + | </ol> | ||
| + | </br> | ||
| + | </br> | ||
| - | < | + | <center><a href="https://2013.igem.org/Team:Penn/MaGellinMotivation">←Previous</a> <a href="https://2013.igem.org/Team:Penn/AssayOverview">Next→</a></center> |
</div> | </div> | ||
</div> | </div> | ||
| + | <div id ="pagefooter"> | ||
| + | <br> | ||
| + | <br> | ||
| + | <center><a href = "https://2013.igem.org/Team:Penn"> Home </a> <a href = "https://static.igem.org/mediawiki/2013/e/e5/Spec_Sheet.pdf" >Spec Sheet</a> <a href = "https://2013.igem.org/Team:Penn/sitemap" >Sitemap</a> | ||
| + | </center> | ||
| + | <br> | ||
| + | Penn iGem © 2013 | ||
| + | </div> | ||
</body> | </body> | ||
Latest revision as of 20:11, 28 October 2013
Toolbox
Constructing a Toolbox
Our goal was to create a toolbox for developing customized site-specific methylases. Like any good toolbox, it includes three components: the tool, the assay, and the automation.- The Tool: a fusion between a DNA binding domain and a methylase enzyme
- The Assay: a digestion based assay, called MaGellin, to measure both methylase activity and sequence specificity
- The Automation: a unique software package that predicts experimental outcomes and analyzes gel electrophoresis images to measure methylase functionality
- Robust – includes functional fusion proteins for comparison
- Standardized – all inclusive in one-plasmid with simple multiple cloning sites, standard bisulfite sequencing primers, and a digestion based methylation assay built in
- Easy to Use – we programmed a software package to predict experimental outcomes and automate gel electrophoresis analysis
- Inexpensive – methylase activity and specificity can be screened by simply digesting and running a gel
- Noiseless – the bacterial chassis has no background CpG methylation
- Open Source – we BioBrick’ed the full plasmid and the methylases
