| − | <!-- # TODO: #6 Fix table caption font--><!-- # TODO: #7 Fix citations links font size--><html lang="en"><head><meta charset="utf-8"/><meta content="width=device-width,initial-scale=1" name="viewport"/><title>Proof Of Concept | iGEM Stockholm</title><script src="https://2020.igem.org/common/MathJax-2.5-latest/MathJax.js?config=TeX-AMS-MML_HTMLorMML"></script><link href="https://2021.igem.org/Template:Stockholm/css/contentCSS?action=raw&ctype=text/css" rel="stylesheet"/></head><body><!-- # TODO: #6 Fix table caption font--><!-- # TODO: #7 Fix citations links font size--><nav class="navbar navbar-expand-xl fixed-top"><div class="container d-flex justify-content-between"><a class="navbar-brand d-lg-inline-block" href="https://2021.igem.org/Team:Stockholm"></a><button aria-controls="navbarNav" aria-expanded="false" aria-label="Toggle navigation" class="navbar-toggler" data-target="#navbarNav" data-toggle="collapse" type="button"><span class="navbar-toggler-icon"></span></button><div class="collapse navbar-collapse" id="navbarNav"><ul class="navbar-nav ml-auto"><li class="nav-item dropdown"><a aria-expanded="false" aria-haspopup="true" class="nav-link dropdown-toggle" data-toggle="dropdown" href="#" id="navbarTeamDropdown" role="button">Team</a><div aria-labelledby="navbarTeamDropdown" class="dropdown-menu"><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Team">Team</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Attributions">Attributions</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Collaborations">Collaborations</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Inclusion">Inclusion</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Sponsors">Sponsors</a></div></li><li class="nav-item dropdown"><a aria-expanded="false" aria-haspopup="true" class="nav-link dropdown-toggle" data-toggle="dropdown" href="#" id="navbarProjectDropdown" role="button">Project</a><div aria-labelledby="navbarProjectDropdown" class="dropdown-menu"><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Contribution">Contribution</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Description">Description</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Design">Design</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Engineering">Engineering</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Experiments">Experiments</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Notebook">Notebook</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Partnership">Partnership</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Proof_Of_Concept">Proof Of Concept</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Results">Results</a></div></li><li class="nav-item dropdown"><a aria-expanded="false" aria-haspopup="true" class="nav-link dropdown-toggle" data-toggle="dropdown" href="#" id="navbarPartsDropdown" role="button">Parts</a><div aria-labelledby="navbarPartsDropdown" class="dropdown-menu"><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Part_Collection">Part Collection</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Parts">Parts</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Model">Model</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Improve">Improve</a></div></li><li class="nav-item dropdown"><a aria-expanded="false" aria-haspopup="true" class="nav-link dropdown-toggle" data-toggle="dropdown" href="#" id="navbarHuman PracticeDropdown" role="button">Human Practice</a><div aria-labelledby="navbarHuman PracticeDropdown" class="dropdown-menu"><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Human_Practices">Human Practices</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Integrated_Human_Practices">Integrated Human Practices</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Entrepreneurship">Entrepreneurship</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Communications">Communications</a></div></li><li class="nav-item"><a class="nav-link" href="https://2021.igem.org/Team:Stockholm/Safety">Safety</a></li></ul></div><div class="d-flex" id="themeSwitchWrapper"><i class="far fa-sun"></i><div id="themeSwitch"><label class="switch" for="themeSwitchInput"><input id="themeSwitchInput" type="checkbox"/><span class="slider round"></span></label></div><i class="far fa-moon"></i></div></div></nav><header class="d-flex justify-content-center align-items-center"><div class="container"><h1>Proof Of Concept</h1><p class="lead pl-1">What actually is our project? Would it work?</p><hr class="my-4"/></div></header><main><div class="container"><div class="row"><div class="sidebar col-lg-3"><div class="nav" id="contents"><h5>Contents</h5><ul></ul></div></div><div class="content col-lg-9"><article><h1>Overview</h1><p>MIKROSKIN aims to be an easy to use rapid test for the detection of dysbiosis in the skin microbiota. To develop our rapid test, we divided the project into smaller parts, to make the development easier and faster, as we were able to work on several parts simultaneously. Our project is divided into three different parts.</p><p>The first part is the aptamer selection, where we use SELEX to select aptamers against the common skin bacteria C.Acnes and one aptamer against all gram-positive skin bacteria. The second part of the project is the analysis of an already discovered aptamer against S.aureus. The third part is the development of the detection method, where we used the colorimetric properties of the polymer PCDA to induce a color change upon aptamer-target binding.</p><p>Our goal for our project was to develop a prototype of our rapid-test, which we could use in presenting our product to potential stakeholders, for the establishment of our business model.</p><h1>Aptamer Selection</h1><h1>Aptamer Testing</h1><p>To develop a functional and highly sensitive rapid test, it is important to have sensitive aptamers. We decided to test the affinity of the aptamers using an affinity assay based on fluorescence detection. The affinity of the aptamers against the immobilized target were measured by detecting emitted fluorescence. Therefore, the aptamers were tagged with a fluorophore on the 5’end.</p><p>The affinity of purchased protein-A aptamer was tested as described. Its affinity to bind protein-A was weak.</p><p>Unfortunately, due to the shortage of time, we were not able to test the binding affinity of our selected aptamers to their targets.</p><h1>Detection Method</h1><p>PCDA polymerisation → Testing of color change → Absorption curves</p><p>Aptamer testing → Binding of aptamer to target → conjugation of aptamer → FACS</p></article><article id="references"><h1>References</h1><ol><li id="citation1"><p class="author">Allen, M. J., & Sheridan, S. C. (2015).</p><cite>Mortality risks during extreme temperature events (ETEs) using a distributed lag non-linear model.</cite><p><span class="journalTitle">International Journal of Biometeorology</span> <span class="journalInfo">62(1), 57-67.</span></p><a class="in-text" href="https://doi.org/10.1007/s00484-015-1117-4" rel="noopener" target="_blank">CrossRef</a><a class="in-text" href="https://scholar.google.com/scholar?q=Mortality risks during extreme temperature events (ETEs) using a distributed lag non-linear model." rel="noopener" target="_blank">Google Scholar</a><a class="in-text" href="#intext1">Back to text</a></li><li id="citation2"><p class="author">Rosano, A., Bella, A., Gesualdo, F., Acampora, A., Pezzotti, P., Marchetti, S., ... & Rizzo, C. (2019).</p><cite>Investigating the impact of influenza on excess mortality in all ages in Italy during recent seasons (2013/14-2016/17 seasons).</cite><p><span class="journalTitle">International Journal of Infectious Diseases</span> <span class="journalInfo">88, 127-134.</span></p><a class="in-text" href="https://doi.org/10.1016/j.ijid.2019.08.003" rel="noopener" target="_blank">CrossRef</a><a class="in-text" href="https://scholar.google.com/scholar?q=Investigating the impact of influenza on excess mortality in all ages in Italy during recent seasons (2013/14-2016/17 seasons)." rel="noopener" target="_blank">Google Scholar</a><a class="in-text" href="#intext2">Back to text</a></li><li id="citation3"><p class="author">Ingalls, B. P. (2013).</p><cite>Mathematical modeling in systems biology: An introduction.</cite><p><span class="details">MIT Press.</span></p><a class="in-text" href="https://books.google.co.in/books?id=OYr6AQAAQBAJ" rel="noopener" target="_blank">Google Books</a><a class="in-text" href="#intext3">Back to text</a></li><li id="citation4"><p class="author"></p><cite>Agriculture: Crop production: Sugarcane. TNAU Agritech Portal.</cite><p><span class="details">(March 15, 2019). Retrieved on June 22, 2020. from </span><a class="in-text" href="https://google.com" rel="noopener" target="_blank">https://google.com</a></p><a class="in-text" href="#intext4">Back to text</a></li><li id="citation5"><p class="author">Author Name. (n.d.).</p><cite>Agriculture: Crop production: Sugarcane. TNAU Agritech Portal.</cite><p><span class="details">Retrieved on June 22, 2020. from </span><a class="in-text" href="https://google.com" rel="noopener" target="_blank">https://google.com</a></p><a class="in-text" href="#intext5">Back to text</a></li></ol></article></div></div></div></main><footer><div class="container"><a class="fafa" href="https://www.facebook.com/igemstockholm/" target="_blank"><i class="fab fa-facebook" style="font-size:60px;"></i></a><a class="fafa" href="https://www.instagram.com/igemstockholm" target="_blank"><i class="fab fa-instagram" style="font-size:60px;"></i></a><a class="fafa" href="https://www.linkedin.com/company/igemstockholm" target="_blank"><i class="fab fa-linkedin" style="font-size:60px;"></i></a><a class="fafa" href="https://www.youtube.com/channel/UCh_a6JvWdh6N_i5tYFFcpyw" target="_blank"><i class="fab fa-youtube" style="font-size:60px;"></i></a><a class="fafa" href="mailto: igem.sthlm@gmail.com" target="_blank"><i class="fas fa-envelope" style="font-size:60px;"></i></a></div><br/><div class="container"><a class="uni-logo" href="https://ki.se" target="_blank"><img src="https://static.igem.org/mediawiki/2021/b/bb/T--Stockholm--img--ki-whiteback-modified.png" style="width:100px;height:100px;"/></a><a class="uni-logo" href="https://kth.se" target="_blank"><img src="https://static.igem.org/mediawiki/2021/e/e2/T--Stockholm--img--kthwhite-modified.png" style="width:100px;height:100px;"/></a><a class="uni-logo" href="https://su.se" target="_blank"><img src="https://static.igem.org/mediawiki/2021/d/da/T--Stockholm--img--stockholmuni-modified.png" style="width:100px;height:100px;"/></a></div><br/><div class="container"><p>Sample template built using the iGEM Wiki Starter Pack by BITS Goa.</p><p>Code released under the MIT license.</p><p>Based on <a href="https://getbootstrap.com">Bootstrap</a> and themes <a href="https://bootswatch.com/flatly/">Flatly</a> and <a href="https://bootswatch.com/darkly/">Darkly</a> from <a href="https://bootswatch.com/">Bootswatch</a>.</p><p>Some content from the <a href="https://2020.igem.org/Team:Example">iGEM Example Wiki</a>. 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| + | <!-- # TODO: #6 Fix table caption font--><!-- # TODO: #7 Fix citations links font size--><html lang="en"><head><meta charset="utf-8"/><meta content="width=device-width,initial-scale=1" name="viewport"/><title>Proof Of Concept | iGEM Stockholm</title><script src="https://2020.igem.org/common/MathJax-2.5-latest/MathJax.js?config=TeX-AMS-MML_HTMLorMML"></script><link href="https://2021.igem.org/Template:Stockholm/css/contentCSS?action=raw&ctype=text/css" rel="stylesheet"/></head><body><!-- # TODO: #6 Fix table caption font--><!-- # TODO: #7 Fix citations links font size--><nav class="navbar navbar-expand-xl fixed-top"><div class="container d-flex justify-content-between"><a class="navbar-brand d-lg-inline-block" href="https://2021.igem.org/Team:Stockholm"></a><button aria-controls="navbarNav" aria-expanded="false" aria-label="Toggle navigation" class="navbar-toggler" data-target="#navbarNav" data-toggle="collapse" type="button"><span class="navbar-toggler-icon"></span></button><div class="collapse navbar-collapse" id="navbarNav"><ul class="navbar-nav ml-auto"><li class="nav-item dropdown"><a aria-expanded="false" aria-haspopup="true" class="nav-link dropdown-toggle" data-toggle="dropdown" href="#" id="navbarTeamDropdown" role="button">Team</a><div aria-labelledby="navbarTeamDropdown" class="dropdown-menu"><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Team">Team</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Attributions">Attributions</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Collaborations">Collaborations</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Inclusion">Inclusion</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Sponsors">Sponsors</a></div></li><li class="nav-item dropdown"><a aria-expanded="false" aria-haspopup="true" class="nav-link dropdown-toggle" data-toggle="dropdown" href="#" id="navbarProjectDropdown" role="button">Project</a><div aria-labelledby="navbarProjectDropdown" class="dropdown-menu"><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Contribution">Contribution</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Description">Description</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Design">Design</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Engineering">Engineering</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Experiments">Experiments</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Notebook">Notebook</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Partnership">Partnership</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Proof_Of_Concept">Proof Of Concept</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Results">Results</a></div></li><li class="nav-item dropdown"><a aria-expanded="false" aria-haspopup="true" class="nav-link dropdown-toggle" data-toggle="dropdown" href="#" id="navbarPartsDropdown" role="button">Parts</a><div aria-labelledby="navbarPartsDropdown" class="dropdown-menu"><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Part_Collection">Part Collection</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Parts">Parts</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Model">Model</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Improve">Improve</a></div></li><li class="nav-item dropdown"><a aria-expanded="false" aria-haspopup="true" class="nav-link dropdown-toggle" data-toggle="dropdown" href="#" id="navbarHuman PracticeDropdown" role="button">Human Practice</a><div aria-labelledby="navbarHuman PracticeDropdown" class="dropdown-menu"><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Human_Practices">Human Practices</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Integrated_Human_Practices">Integrated Human Practices</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Entrepreneurship">Entrepreneurship</a><a class="dropdown-item" href="https://2021.igem.org/Team:Stockholm/Communication">Communication</a></div></li><li class="nav-item"><a class="nav-link" href="https://2021.igem.org/Team:Stockholm/Safety">Safety</a></li></ul></div><div class="d-flex" id="themeSwitchWrapper"><i class="far fa-sun"></i><div id="themeSwitch"><label class="switch" for="themeSwitchInput"><input id="themeSwitchInput" type="checkbox"/><span class="slider round"></span></label></div><i class="far fa-moon"></i></div></div></nav><header class="d-flex justify-content-center align-items-center"><div class="container"><h1>Proof Of Concept</h1><p class="lead pl-1">What actually is our project? Would it work?</p><hr class="my-4"/></div></header><main><div class="container"><div class="row"><div class="sidebar col-lg-3"><div class="nav" id="contents"><h5>Contents</h5><ul></ul></div></div><div class="content col-lg-9"><article><h1>Overview</h1><p>MIKROSKIN aims to be an easy to use rapid test for the detection of dysbiosis in the skin microbiota. To develop our rapid test, we divided the project into smaller parts, to make the development easier and faster, as we were able to work on several parts simultaneously. Our project is divided into three different parts.</p><p>The first part is the aptamer selection, where we use SELEX to select aptamers against the common skin bacteria C.Acnes and one aptamer against all gram-positive skin bacteria. The second part of the project is the analysis of an already discovered aptamer against S.aureus. The third part is the development of the detection method, where we used the colorimetric properties of the polymer PCDA to induce a color change upon aptamer-target binding.</p><p>Our goal for our project was to develop a prototype of our rapid-test, which we could use in presenting our product to potential stakeholders, for the establishment of our business model.</p><h1>Aptamer Selection</h1><h1>Aptamer Testing</h1><p>To develop a functional and highly sensitive rapid test, it is important to have sensitive aptamers. We decided to test the affinity of the aptamers using an affinity assay based on fluorescence detection. The affinity of the aptamers against the immobilized target were measured by detecting emitted fluorescence. Therefore, the aptamers were tagged with a fluorophore on the 5’end.</p><p>The affinity of purchased protein-A aptamer was tested as described. Its affinity to bind protein-A was weak.</p><p>Unfortunately, due to the shortage of time, we were not able to test the binding affinity of our selected aptamers to their targets.</p><h1>Detection Method</h1><p>PCDA polymerisation → Testing of color change → Absorption curves</p><p>Aptamer testing → Binding of aptamer to target → conjugation of aptamer → FACS</p></article><article id="references"><h1>References</h1><ol><li id="citation1"><p class="author">Allen, M. J., & Sheridan, S. C. (2015).</p><cite>Mortality risks during extreme temperature events (ETEs) using a distributed lag non-linear model.</cite><p><span class="journalTitle">International Journal of Biometeorology</span> <span class="journalInfo">62(1), 57-67.</span></p><a class="in-text" href="https://doi.org/10.1007/s00484-015-1117-4" rel="noopener" target="_blank">CrossRef</a><a class="in-text" href="https://scholar.google.com/scholar?q=Mortality risks during extreme temperature events (ETEs) using a distributed lag non-linear model." rel="noopener" target="_blank">Google Scholar</a><a class="in-text" href="#intext1">Back to text</a></li><li id="citation2"><p class="author">Rosano, A., Bella, A., Gesualdo, F., Acampora, A., Pezzotti, P., Marchetti, S., ... & Rizzo, C. (2019).</p><cite>Investigating the impact of influenza on excess mortality in all ages in Italy during recent seasons (2013/14-2016/17 seasons).</cite><p><span class="journalTitle">International Journal of Infectious Diseases</span> <span class="journalInfo">88, 127-134.</span></p><a class="in-text" href="https://doi.org/10.1016/j.ijid.2019.08.003" rel="noopener" target="_blank">CrossRef</a><a class="in-text" href="https://scholar.google.com/scholar?q=Investigating the impact of influenza on excess mortality in all ages in Italy during recent seasons (2013/14-2016/17 seasons)." rel="noopener" target="_blank">Google Scholar</a><a class="in-text" href="#intext2">Back to text</a></li><li id="citation3"><p class="author">Ingalls, B. P. (2013).</p><cite>Mathematical modeling in systems biology: An introduction.</cite><p><span class="details">MIT Press.</span></p><a class="in-text" href="https://books.google.co.in/books?id=OYr6AQAAQBAJ" rel="noopener" target="_blank">Google Books</a><a class="in-text" href="#intext3">Back to text</a></li><li id="citation4"><p class="author"></p><cite>Agriculture: Crop production: Sugarcane. TNAU Agritech Portal.</cite><p><span class="details">(March 15, 2019). Retrieved on June 22, 2020. from </span><a class="in-text" href="https://google.com" rel="noopener" target="_blank">https://google.com</a></p><a class="in-text" href="#intext4">Back to text</a></li><li id="citation5"><p class="author">Author Name. (n.d.).</p><cite>Agriculture: Crop production: Sugarcane. TNAU Agritech Portal.</cite><p><span class="details">Retrieved on June 22, 2020. from </span><a class="in-text" href="https://google.com" rel="noopener" target="_blank">https://google.com</a></p><a class="in-text" href="#intext5">Back to text</a></li></ol></article></div></div></div></main><footer><div class="container"><a class="fafa" href="https://www.facebook.com/igemstockholm/" target="_blank"><i class="fab fa-facebook" style="font-size:60px;"></i></a><a class="fafa" href="https://www.instagram.com/igemstockholm" target="_blank"><i class="fab fa-instagram" style="font-size:60px;"></i></a><a class="fafa" href="https://www.linkedin.com/company/igemstockholm" target="_blank"><i class="fab fa-linkedin" style="font-size:60px;"></i></a><a class="fafa" href="https://www.youtube.com/channel/UCh_a6JvWdh6N_i5tYFFcpyw" target="_blank"><i class="fab fa-youtube" style="font-size:60px;"></i></a><a class="fafa" href="mailto: igem.sthlm@gmail.com" target="_blank"><i class="fas fa-envelope" style="font-size:60px;"></i></a></div><br/><div class="container"><a class="uni-logo" href="https://ki.se" target="_blank"><img src="https://static.igem.org/mediawiki/2021/b/bb/T--Stockholm--img--ki-whiteback-modified.png" style="width:100px;height:100px;"/></a><a class="uni-logo" href="https://kth.se" target="_blank"><img src="https://static.igem.org/mediawiki/2021/e/e2/T--Stockholm--img--kthwhite-modified.png" style="width:100px;height:100px;"/></a><a class="uni-logo" href="https://su.se" target="_blank"><img src="https://static.igem.org/mediawiki/2021/d/da/T--Stockholm--img--stockholmuni-modified.png" style="width:100px;height:100px;"/></a></div><br/><div class="container"><p>Sample template built using the iGEM Wiki Starter Pack by BITS Goa.</p><p>Code released under the MIT license.</p><p>Based on <a href="https://getbootstrap.com">Bootstrap</a> and themes <a href="https://bootswatch.com/flatly/">Flatly</a> and <a href="https://bootswatch.com/darkly/">Darkly</a> from <a href="https://bootswatch.com/">Bootswatch</a>.</p><p>Some content from the <a href="https://2020.igem.org/Team:Example">iGEM Example Wiki</a>. 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