Difference between revisions of "Team:NAIT Edmonton/Trial11"

 
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<center><div class="top_slogan">The Project</div></center>
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<div class="flexslider">
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    <ul class="slides">
 +
    <li><a href=""><img src="https://static.igem.org/mediawiki/2015/6/62/NAIT_Slide_Pedway.jpg" alt="" title="" border="0"/></a>
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            <div class="flex-caption">
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                <h2></h2>
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                    <p><strong><a href=""></a></strong></p>
 +
                </div>
 +
            </li>
 +
            <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Desc"><img src="https://static.igem.org/mediawiki/2015/5/56/NAIT_Slide_1_Project.jpg"alt="" title="" border="0"/></a>
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                <h2></h2>
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                    <p></p>
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            </li>
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            <li><a href="https://2015.igem.org/Team:NAIT_Edmonton/Practices"><img src="https://static.igem.org/mediawiki/2015/d/db/NAIT_Slide_2_Whiteboard.jpg" alt="" title="" border="0"/></a>
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                <h2></h2>
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                    <p></p>
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                <h2></h2>
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<div id="wrap5">
  
<h2>Sodium Dodecyl Sulphide Polyacrylamide Gel Electrophoresis (SDS PAGE) is one of the most prominent techniques used in protein research. However, one of its limitations, more specifically the loss of an accurate point of reference post staining, is attributed to inefficiency and the wasting of time during the research process. From past observations, some proteins inherently produce certain colours post-silver staining. We hypothesized that certain configurations of amino acids produces specific colours when reacting with the reagents of silver staining. In the hopes of creating a new molecular weight ladder, our team designed sequences to code for novel proteins that produce colour. Coloured proteins can also pave the way for new types of colorimetric assays and a less expensive and more accessible counterpart to antibody tags in the future.</h2> <br>
+
  <div class="section_one_three">
 +
        <h2 class="centered_title">Achievements</h2>
 +
        <div class="roundimg"><a href="https://2015.igem.org/Team:NAIT_Edmonton/Achievements" title=""><img src="https://static.igem.org/mediawiki/2015/5/50/NAIT_Medal_Icon.png" width="125px" alt="" title="" /></a></div>
 +
        <p class="centered_text">
 +
Despite the time constraints, our team achieved a lot this summer and we are proud of what we have accomplished.
 +
        </p>
 +
        <a href="https://2015.igem.org/Team:NAIT_Edmonton/Achievements" class="more">read more</a>
 +
        </div>
 +
       
 +
  <div class="section_one_three">
 +
        <h2 class="centered_title">iGEM 2015</h2>
 +
        <div class="roundimg"><a href="https://2015.igem.org" title=""><img src="https://static.igem.org/mediawiki/2015/0/05/NAIT_Icon_services.png" alt="" title="" /></a></div>
 +
        <p class="centered_text">
 +
Learn about the exciting opportunities the iGEM foundation has to offer! The experience was incredible!
 +
        </p>
 +
        <a href="https://2015.igem.org" class="more">read more</a>
 +
        </div>
  
<div class="accordion">
+
  <div class="section_one_three">
<div class="accordion-section">
+
        <h2 class="centered_title">Contact Us!</h2>
<a class="accordion-section-title" href="#accordion-1">Background</a>
+
        <div class="roundimg"><a href="https://2015.igem.org/Team:NAIT_Edmonton/ContactUs" title=""><img src="https://static.igem.org/mediawiki/2015/2/29/NAIT_Icon_contact.png" alt="" title="" /></a></div>
<div id="accordion-1" class="accordion-section-content">
+
        <p class="centered_text">
The structural and functional study of the proteins expressed by a genome is
+
Please contact our team if you want more information about our project or have any other questions or concerns.
 +
        </p>
 +
        <a href="https://2015.igem.org/Team:NAIT_Edmonton/ContactUs" class="more">read more</a>
 +
        </div>
 +
 
 +
</div>
  
called proteomics. This relatively novel science uses different methodologies in order to
 
  
separate and identify specific proteins of interest. Among these techniques, SDS-PAGE
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  <div class="clear"></div>
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</div>
  
plays an essential role due to its high sensitivity, low sample volume requirement, and
 
  
high popularity. Negatively charged proteins migrate towards the positive electrode
 
 
according to their size and charge. Smaller proteins migrate further in a given amount of
 
 
time. As proteins are separated in this manner, users load molecular weight standards
 
 
to estimate the size (in kDa) of the proteins present in their sample. Once the proteins of
 
 
a single sample have been isolated and are embedded in the polyacrylamide (PA) gel
 
 
matrix, staining procedures are used to visualize them.</p>
 
 
<br>
 
 
<center><img src="http://upload.wikimedia.org/wikipedia/commons/4/46/SDS-PAGE_Electrophoresis.png" width="750px"></center>
 
 
<br>
 
 
<p style="font-size:15;">Organic dyes, such as Coomassie blue, can be used for this purpose;
 
 
nevertheless, their low sensitivity and a detection range that goes from 1 to 50 ng can
 
 
be a challenge for detecting low abundance proteins (Jin, Huang, Yoo, & Choi, 2006). A
 
 
higher sensitivity can be achieved by fluorescent staining techniques (from 0.1 to 10
 
 
ng.); however, UV instruments are necessary in order to read the data (Jin et al., 2006).
 
 
The most sensitive method up to date is radiolabeling, but the requirement of hazardous
 
 
isotopes and their complex management makes it a complicated procedure (Jin et al.,
 
 
2006). Silver staining is a method that offers great sensitivity and an easy to handle
 
 
protocol, thus making it one of the most commonly used staining methods. </p>
 
 
<br>
 
 
<center><img src="http://www.bio-rad.com/webroot/web/images/lsr/products/electrophoresis/product_overlay_content/global/lsr_biosafe_coomasie_gel.jpg"></center>
 
 
</div><!--end .accordion-section-content-->
 
</div><!--end .accordion-section-->
 
 
<div class="accordion-section">
 
<a class="accordion-section-title" href="#accordion-2">The Problem</a>
 
<div id="accordion-2" class="accordion-section-content">
 
<p>Difficulties with silver staining arise when the molecular weight markers are re-
 
 
colored golden-brown in the staining process. Markers offer evenly distributed proteins
 
 
that show bands of equal intensity and known size. Researchers can compare these
 
 
bands with their sample and identify the protein they are looking for based on its size. A
 
 
subset of these markers has color-coded standard proteins to facilitate the identification
 
 
of each band. Post-silver staining, the users lose the ability to use the color code as a
 
 
reference.</p>
 
 
<br>
 
 
<center><img src="http://labs.mmg.pitt.edu/gjoerup/silver_stain.jpg" width="750px"></center>
 
 
</div><!--end .accordion-section-content-->
 
</div><!--end .accordion-section-->
 
 
<div class="accordion-section">
 
<a class="accordion-section-title" href="#accordion-3">Our Goal</a>
 
<div id="accordion-3" class="accordion-section-content">
 
<p>Our goal is to develop a marker that, when interacting with the reagents used in
 
 
the staining protocol, will develop colour bands in specific positions so as to help in the
 
 
identification of the protein(s) of interest post-staining. In order to do so, investigation of
 
 
how specific amino acids react with silver staining reagents is underway by our team.
 
 
This will have as an outcome the creation of novel proteins that contain an excess of a
 
 
particular amino acid and/or chemical modifications that will generate a specific colour
 
 
after treating it with silver staining reagents. To obtain such proteins, the introduction of
 
 
novel nucleotide sequences into a plasmid would be done first by in vitro transcription
 
 
translation and later by transforming E. coli cells with expression vectors.</p>
 
 
</div><!--end .accordion-section-content-->
 
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                          _ __  _
 
                        ;'  '',)
 
                        /;6 , ;/
 
                        (Y)_:., |
 
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                          |;  ,.:\
 
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        eduardo  |',:| \_ \ |_|;\_
 
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Latest revision as of 20:44, 6 August 2015

Team NAIT 2015