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| − | {{NAIT_Edmonton/CSS}}
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| − | <html>
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| − | <head>
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| − | <title>Team NAIT 2015</title>
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| − | <link href='http://fonts.googleapis.com/css?family=Source+Sans+Pro' rel='stylesheet' type='text/css'>
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| − | <script type="text/javascript" src="https://ajax.googleapis.com/ajax/libs/jquery/1.6.2/jquery.min.js"></script>
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| − | <script type="text/javascript" src="https://2015.igem.org/index.php?title=
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| − | Template:NAIT_Edmonton/js/jquery.flexslider"></script>
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| − | <script type="text/javascript" charset="utf-8">
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| − | var $ = jQuery.noConflict();
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| − | $(window).load(function() {
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| − | $('.flexslider').flexslider({
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| − | animation: "slide"
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| − | });
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| − | });
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| − | </script>
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| − | </head>
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| − | <body>
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| − |
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| − | <style type="text/css">
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| − | .footer{position:fixed;bottom:0px}
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| − | </style>
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| − |
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| − | <div id="header">
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| − | <div class="header_content">
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| − | <div class="logo"><a href="https://2015.igem.org/Team:NAIT_Edmonton"><img src="https://static.igem.org/mediawiki/2015/4/4a/NAIT_Logo_joy.png" width="141" height="140"></a></div>
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| − |
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| − | <div class="menu">
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| − | <ul>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton">home</a>
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| − | </li>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Team">team</a>
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| − | <ul>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Bios">bios</a></li>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Attributions">attributions</a></li>
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| − | <li><a href="https://igem.org/Team.cgi">official team profile</a>
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| − | </ul>
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| − | </li>
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| − | <li class="selected"><a href="https://2015.igem.org/Team:NAIT_Edmonton/Project">project</a>
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| − | <ul>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Desc">description</a></li>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Protocols">experiment and protcols</a></li>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Results">parts and results</a></li>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Modeling">modeling</a></li>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Software">software</a></li>
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| − | </ul>
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| − | </li>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Human">human practices</a>
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| − | <ul>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Outreach">community outreach</a></li>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Practices">policy and practices</a></li>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Collaborations">collaborations</a></li>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Entrepreunership">entrepreunership</a></li>
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| − | </ul>
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| − | </li>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Notebook">notebook</a>
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| − | <ul>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Safety">lab safety</a></li>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Measurement">measurement</a></li>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Logbook">log book</a></li>
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| − | </ul>
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| − | </li>
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| − | </ul>
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| − | </div>
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| − | </div>
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| − | </div>
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| − | <div id="wrap">
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| − | <center><div class="top_slogan">The Project</div></center>
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| − | <div id="nav_content">
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| − | <h1>Background</h1>
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| − | <p>The structural and functional study of the proteins expressed by a genome is
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| − | called proteomics. This relatively novel science uses different methodologies in order to
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| − | separate and identify specific proteins of interest. Among these techniques, SDS-PAGE
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| − | plays an essential role due to its high sensitivity, low sample volume requirement, and
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| − | high popularity. Negatively charged proteins migrate towards the positive electrode
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| − | according to their size and charge. Smaller proteins migrate further in a given amount of
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| − | time. As proteins are separated in this manner, users load molecular weight standards
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| − | to estimate the size (in kDa) of the proteins present in their sample. Once the proteins of
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| − | a single sample have been isolated and are embedded in the polyacrylamide (PA) gel
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| − | matrix, staining procedures are used to visualize them.</p>
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| − | <br>
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| − | <center><img src="http://upload.wikimedia.org/wikipedia/commons/4/46/SDS-PAGE_Electrophoresis.png" width="750px"></center>
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| − |
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| − | <br>
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| − | <p>Organic dyes, such as Coomassie blue, can be used for this purpose;
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| − | nevertheless, their low sensitivity and a detection range that goes from 1 to 50 ng can
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| − | be a challenge for detecting low abundance proteins (Jin, Huang, Yoo, & Choi, 2006). A
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| − | higher sensitivity can be achieved by fluorescent staining techniques (from 0.1 to 10
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| − | ng.); however, UV instruments are necessary in order to read the data (Jin et al., 2006).
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| − | The most sensitive method up to date is radiolabeling, but the requirement of hazardous
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| − | isotopes and their complex management makes it a complicated procedure (Jin et al.,
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| − | 2006). Silver staining is a method that offers great sensitivity and an easy to handle
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| − | protocol, thus making it one of the most commonly used staining methods. </p>
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| − | <br>
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| − | <center><img src="http://www.bio-rad.com/webroot/web/images/lsr/products/electrophoresis/product_overlay_content/global/lsr_biosafe_coomasie_gel.jpg"></center>
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| − | <br><br>
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| − | <h1>The Problem </h1>
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| − | <p>Difficulties with silver staining arise when the molecular weight markers are re-
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| − | colored golden-brown in the staining process. Markers offer evenly distributed proteins
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| − | that show bands of equal intensity and known size. Researchers can compare these
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| − | bands with their sample and identify the protein they are looking for based on its size. A
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| − | subset of these markers has color-coded standard proteins to facilitate the identification
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| − | of each band. Post-silver staining, the users lose the ability to use the color code as a
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| − | reference.</p>
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| − | <br>
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| − | <center><img src="http://labs.mmg.pitt.edu/gjoerup/silver_stain.jpg"></center>
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| − | <br><br>
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| − | <h1>Our Goal</h1>
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| − | <p>Our goal is to develop a marker that, when interacting with the reagents used in
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| − | the staining protocol, will develop colour bands in specific positions so as to help in the
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| − | identification of the protein(s) of interest post-staining. In order to do so, investigation of
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| − | how specific amino acids react with silver staining reagents is underway by our team.
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| − | This will have as an outcome the creation of novel proteins that contain an excess of a
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| − | particular amino acid and/or chemical modifications that will generate a specific colour
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| − | after treating it with silver staining reagents. To obtain such proteins, the introduction of
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| − | novel nucleotide sequences into a plasmid would be done first by in vitro transcription
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| − | translation and later by transforming E. coli cells with expression vectors.</p>
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| − | <br>
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| − | </div>
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| − | <div id="nav_bar">
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| − | <li><a href="/">Background</a></li>
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| − | <li><a href="/">The Problem</a></li>
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| − | <li><a href="/">Our Goal</a></li>
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| − | </div>
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| − | </div>
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| − | <div class="clear"></div>
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| − | </div>
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| − | <div class="footer">
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| − | <div class="footer_content">
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| − | <div class="footer_left">
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| − | <ul class="footer_menu">
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton">home</a></li>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Team">team</a></li>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Project">project</a></li>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Human">human practices</a></li>
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| − | <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Notebook">notebook</a></li>
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| − | </ul>
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| − | </div>
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| − | <div class="footer_right">
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| − | <ul class="social_icons">
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| − | <li><a href="http://www.facebook.com/pages/IGEM-2015-NAIT-Edmonton/884025281659292"><img src="https://static.igem.org/mediawiki/2015/a/a3/NAIT_Icon_facebook.png" alt="" title="" /></a></li>
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| − | <li><a href="http://www.twitter.com/TeamNAIT2015"><img src="https://static.igem.org/mediawiki/2015/6/64/NAIT_Icon_twitter.png" alt="" title="" /></a></li>
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| − | </ul>
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| − | </div>
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| − | <div class="clear"></div>
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| − | </div>
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| − | </div>
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| − | </body>
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| − | </html>
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