Team:Penn/MaGellinToolbox

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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.
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<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.
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<li>The Tool: a fusion between a DNA binding domain and a methylase enzyme</li>
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<li>The Assay: a digestion based assay, called MaGellin, to measure both methylase activity and sequence specificity</li>
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<li>The Automation: a unique software package that predicts experimental outcomes and analyzes gel electrophoresis images to measure methylase functionality</li>
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<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>
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Our toolbox satisfies 6 requirements:
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<li>Robust – includes functional fusion proteins for comparison</li>
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<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>
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<li>Easy to Use – we programmed a software package to predict experimental outcomes and automate gel electrophoresis analysis</li>
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<li>Inexpensive – methylase activity and specificity can be screened by simply digesting and running a gel</li>
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<li>Noiseless – the bacterial chassis has no background CpG methylation</li>
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<li>Open Source – we<a href="https://2013.igem.org/Team:Penn/Biobricks"> BioBrick’ed</a> the full plasmid and the methylases</li>
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<center><a href="https://2013.igem.org/Team:Penn/MaGellinMotivation">&#8592;Previous</a> <a href="https://2013.igem.org/Team:Penn/AssayOverview">Next&#8594;</a></center>
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<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>
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Penn iGem &copy; 2013
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Latest revision as of 20:11, 28 October 2013

Penn iGEM

Toolbox




For a detailed, graphical explanation of the MaGellin work flow, please download the MaGellin Workflow Specifications Sheet, which includes all of the steps in the MaGellin workflow.


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.
  1. The Tool: a fusion between a DNA binding domain and a methylase enzyme
  2. The Assay: a digestion based assay, called MaGellin, to measure both methylase activity and sequence specificity
  3. The Automation: a unique software package that predicts experimental outcomes and analyzes gel electrophoresis images to measure methylase functionality


Toolbox
Toolbox with the three essential components.



Our toolbox satisfies 6 requirements:
  1. Robust – includes functional fusion proteins for comparison
  2. Standardized – all inclusive in one-plasmid with simple multiple cloning sites, standard bisulfite sequencing primers, and a digestion based methylation assay built in
  3. Easy to Use – we programmed a software package to predict experimental outcomes and automate gel electrophoresis analysis
  4. Inexpensive – methylase activity and specificity can be screened by simply digesting and running a gel
  5. Noiseless – the bacterial chassis has no background CpG methylation
  6. Open Source – we BioBrick’ed the full plasmid and the methylases


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