Difference between revisions of "Team:Tianjin/Measurement"

 
(23 intermediate revisions by 4 users not shown)
Line 8: Line 8:
 
         <meta content="width=device-width,initial-scale=1.0,maximum-scale=1.0,user-scalable=0;" name="viewport">
 
         <meta content="width=device-width,initial-scale=1.0,maximum-scale=1.0,user-scalable=0;" name="viewport">
 
         <meta name="renderer" content="webkit">
 
         <meta name="renderer" content="webkit">
         <meta name="author" content="773715181 HanJiaxiao">
+
         <meta name="author" content="773715181 Jiaxiao Han">
 
         <meta content="2018iGEM Team:Tianjin Kai System to control biological clock" name="description">
 
         <meta content="2018iGEM Team:Tianjin Kai System to control biological clock" name="description">
 
         <meta content="Team:Tianjin,iGEM:Tianjin,iGEM,2018iGEM" name="keywords">
 
         <meta content="Team:Tianjin,iGEM:Tianjin,iGEM,2018iGEM" name="keywords">
Line 15: Line 15:
 
         <link rel="stylesheet" type="text/css" href="https://2018.igem.org/Team:Tianjin/css/bootstrap?action=raw&ctype=text/css">
 
         <link rel="stylesheet" type="text/css" href="https://2018.igem.org/Team:Tianjin/css/bootstrap?action=raw&ctype=text/css">
 
         <link rel="stylesheet" type="text/css" href="https://2018.igem.org/Team:Tianjin/css/base?action=raw&ctype=text/css">
 
         <link rel="stylesheet" type="text/css" href="https://2018.igem.org/Team:Tianjin/css/base?action=raw&ctype=text/css">
         <link href="//netdna.bootstrapcdn.com/font-awesome/4.7.0/css/font-awesome.min.css" rel="stylesheet">
+
         <link rel="stylesheet" type="text/css" href="https://2018.igem.org/Team:Tianjin/css/font-awesome?action=raw&ctype=text/css">
 
         <link rel="stylesheet" type="text/css" href="https://2018.igem.org/Team:Tianjin/css/text?action=raw&ctype=text/css">
 
         <link rel="stylesheet" type="text/css" href="https://2018.igem.org/Team:Tianjin/css/text?action=raw&ctype=text/css">
 
         <script src="https://2018.igem.org/Team:Tianjin/js/bootstrap?action=raw&ctype=text/javascript"></script>
 
         <script src="https://2018.igem.org/Team:Tianjin/js/bootstrap?action=raw&ctype=text/javascript"></script>
 +
        <link rel="stylesheet" type="text/css" href="https://2018.igem.org/Team:Tianjin/css/slides?action=raw&ctype=text/css">
 
</head>
 
</head>
 
<!-- delete.css  -->
 
<!-- delete.css  -->
Line 28: Line 29:
 
.igem_2018_team_menu{display:none !important;}
 
.igem_2018_team_menu{display:none !important;}
 
</style>
 
</style>
<script src="https://use.fontawesome.com/597ba5ca72.js"></script>
+
 
 
<script>
 
<script>
 
$('#top_title').remove();$("#globalWrapper").removeAttr("id");$("#content").removeAttr("id");$("#HQ_page").removeAttr("id");$('.mw-body').removeClass('mw-body');$('.igem_2018_team_mobile_bar').remove();
 
$('#top_title').remove();$("#globalWrapper").removeAttr("id");$("#content").removeAttr("id");$("#HQ_page").removeAttr("id");$('.mw-body').removeClass('mw-body');$('.igem_2018_team_mobile_bar').remove();
Line 62: Line 63:
 
                 </a>
 
                 </a>
 
                 <ul class="dropdown-menu" style="left: -14px;">
 
                 <ul class="dropdown-menu" style="left: -14px;">
                     <li><a href="https://2018.igem.org/Team:Tianjin/Description">DESCRIPTION</a></li>
+
                     <li><a href="https://2018.igem.org/Team:Tianjin/Description">BACKGROUND</a></li>
 
                     <li><a href="https://2018.igem.org/Team:Tianjin/Design">DESIGN</a></li>
 
                     <li><a href="https://2018.igem.org/Team:Tianjin/Design">DESIGN</a></li>
 
                     <li><a href="https://2018.igem.org/Team:Tianjin/Experiments">EXPERIMENTS</a></li>
 
                     <li><a href="https://2018.igem.org/Team:Tianjin/Experiments">EXPERIMENTS</a></li>
Line 95: Line 96:
 
                 </a>
 
                 </a>
 
                 <ul class="dropdown-menu">
 
                 <ul class="dropdown-menu">
                     <li><a href="https://2018.igem.org/Team:Tianjin/Human_Practices">INTEGRATED HP</a></li>
+
                     <li><a href="https://2018.igem.org/Team:Tianjin/Human_Practices">INTEGRATED</a></li>
 
                     <li><a href="https://2018.igem.org/Team:Tianjin/Collaborations">COLLABORATION</a></li>
 
                     <li><a href="https://2018.igem.org/Team:Tianjin/Collaborations">COLLABORATION</a></li>
                     <li><a href="https://2018.igem.org/Team:Tianjin/Public_Engagement">EDU&PUBLIC ENGAGEMENT</a></li>
+
                     <li><a href="https://2018.igem.org/Team:Tianjin/Public_Engagement">PUBLIC ENGAGEMENT</a></li>
 
                 </ul>
 
                 </ul>
 
             </li>
 
             </li>
Line 111: Line 112:
 
             </li>
 
             </li>
 
             <li>
 
             <li>
                 <a href="https://igem.org/2018_Judging_Form?team=Tianjin">
+
                 <a href="https://2018.igem.org/Team:Tianjin/Judging">
                     JUDGING FORM                
+
                     FOR JUDGES                
 
                 </a>
 
                 </a>
 
             </li>
 
             </li>
Line 120: Line 121:
 
     </nav>
 
     </nav>
 
     <script type="text/javascript" src="https://2018.igem.org/Team:Tianjin/js/navigation?action=raw&ctype=text/javascript"></script>
 
     <script type="text/javascript" src="https://2018.igem.org/Team:Tianjin/js/navigation?action=raw&ctype=text/javascript"></script>
     <div id="fill" style="width: 100%;margin-top:-14px;"></div>
+
      
 
+
    <div id="MainPic">
+
        <div id="bannercontent">
+
            <div id="BgChange">
+
                <img src="https://static.igem.org/mediawiki/2018/7/72/T--Tianjin--partB1.jpg" style="width: 100%;">
+
            </div>
+
        </div>
+
    </div>
+
 
     <div class="container ">
 
     <div class="container ">
 
         <div class="row">
 
         <div class="row">
 
             <div class="col-xs-12 titleBox">
 
             <div class="col-xs-12 titleBox">
 
                 <div class="title title-big">
 
                 <div class="title title-big">
                     <p>Measurment</p>
+
                     <p>MEASUREMENT</p>
 
                 </div>
 
                 </div>
 
             </div>
 
             </div>
Line 144: Line 137:
 
                 <p>
 
                 <p>
 
                     In pursuit of a rigorously scientific result, we have adopted multiple measuring approaches throughout the whole project, among which two predominant methods described in extreme detail in wiki, would inspire relative researchers.<br><br>
 
                     In pursuit of a rigorously scientific result, we have adopted multiple measuring approaches throughout the whole project, among which two predominant methods described in extreme detail in wiki, would inspire relative researchers.<br><br>
                     As two sophisticated and renowned reporters in biotechnological domain, fluorescent protein and luciferase were opted to illustrate the effectiveness of our constructed oscillating system, detected by combined means and instrument. Mcherry and EYFP, two selected fluorescent reporters were measured by both fluorescence spectrophotometer and Leica DMi8 respectively. Leica DMi8 allows us to visually and intuitively see the expression of the fluorescent protein EYFP in Saccharomyces cerevisiae. In terms of luciferase, in addition to commonly used firefly luciferase, a courageous attempt we made was to choose a novel luciferase Nanoluc and its live cell substrate in spite of no records of the usefulness in Saccharomyces cerevisiae before and obtained expected outcome successfully.  
+
                     As two sophisticated and renowned reporters in biotechnological domain, fluorescent protein and luciferase were opted to illustrate the effectiveness of our constructed oscillating system, detected by combined means and instrument. Mcherry and EYFP, two selected fluorescent reporters were measured by both fluorescence spectrophotometer and Leica DMi8 respectively. Leica DMi8 allows us to visually and intuitively see the expression of the fluorescent protein EYFP in <em>Saccharomyces cerevisiae</em> In terms of luciferase, in addition to commonly used firefly luciferase, a courageous attempt we made was to choose a novel luciferase NanoLuc and its live cell substrate in spite of no records of the usefulness in <em>Saccharomyces cerevisiae</em> before and obtained expected outcome successfully.  
 
                 </p>
 
                 </p>
 
             </div>
 
             </div>
Line 163: Line 156:
 
                                 <div class="col-xs-12 text">
 
                                 <div class="col-xs-12 text">
 
                                     <p>
 
                                     <p>
                                       To achieve the best measurement of our oscillating system, we chose two different kinds of reporter gene as our downstream reporter, fluorescent protein gene and luciferase gene,  which are both widely used in the biotechnological labs.
+
                                       To achieve the best measurement of our oscillatory system, we chose two different kinds of reporter gene as our downstream reporters, fluorescent protein gene and luciferase gene,  which are both widely used in the biotechnological labs.
 
                                     </p>
 
                                     </p>
 
                                 </div>
 
                                 </div>
Line 188: Line 181:
 
                                     </table>
 
                                     </table>
 
                                 </div>
 
                                 </div>
                                 <div class="col-lg-12 picture">
+
                                 <div class="col-md-12 col-xs-12">
 +
                                    <div class="wrap">
 +
                                        <div class="photo-container">
 +
                                            <div class="display-left" onclick="" style="right:0px">
 +
                                                <a>&#10094;</a>
 +
                                            </div>
 +
                                            <div class="Slides">
 +
                                                <div class="mySlides">
 +
                                                    <img src="https://static.igem.org/mediawiki/2018/6/61/T--Tianjin--comic1.1.gif">     
 +
                                                </div>
 +
                                                <div class="mySlides">
 +
                                                    <img src="https://static.igem.org/mediawiki/2018/3/30/T--Tianjin--comic1.2.gif" >         
 +
                                                </div>
 +
                                                <div class="mySlides">
 +
                                                    <img src="https://static.igem.org/mediawiki/2018/8/89/T--Tianjin--comic1.3.gif" >         
 +
                                                </div>
 +
                                                <div class="mySlides">
 +
                                                    <img src="https://static.igem.org/mediawiki/2018/c/cc/T--Tianjin--comic1.4.gif" >         
 +
                                                </div>
 +
                                                <div class="mySlides">
 +
                                                    <img src="https://static.igem.org/mediawiki/2018/5/59/T--Tianjin--comic1.5.gif" >         
 +
                                                </div>             
 +
                                                <div class="mySlides">
 +
                                                    <img src="https://static.igem.org/mediawiki/2018/4/49/T--Tianjin--comic1.6.gif" >
 +
                                                </div>
 +
                                                <div class="mySlides">
 +
                                                    <img src="https://static.igem.org/mediawiki/2018/7/7a/T--Tianjin--comic1.7.gif" >
 +
                                                </div>
 +
                                                <div class="mySlides">
 +
                                                    <img src="https://static.igem.org/mediawiki/2018/8/86/T--Tianjin--comic1.8.gif" >
 +
                                                </div>
 +
                                                <div class="mySlides">
 +
                                                    <img src="https://static.igem.org/mediawiki/2018/2/2c/T--Tianjin--comic1.9.gif" >
 +
                                                </div>
 +
                                                <div class="mySlides">
 +
                                                    <img src="https://static.igem.org/mediawiki/2018/e/e2/T--Tianjin--comic1.10.gif" >
 +
                                                </div>
 +
                                            </div>
 +
                                            <div class="display-right" onclick="" style="left:0px;">
 +
                                                <a>&#10095;</a>
 +
                                            </div>
 +
                                        </div>
 +
                                        <div class="control">
 +
                                            <div class="cr"><a><i class="fa fa-circle" aria-hidden="true"></i></a></div>
 +
                                            <div class="cr"><a><i class="fa fa-circle" aria-hidden="true"></i></a></div>
 +
                                            <div class="cr"><a><i class="fa fa-circle" aria-hidden="true"></i></a></div>
 +
                                            <div class="cr"><a ><i class="fa fa-circle" aria-hidden="true"></i></a></div>
 +
                                            <div class="cr"><a ><i class="fa fa-circle" aria-hidden="true"></i></a></div>
 +
                                            <div class="cr"><a ><i class="fa fa-circle" aria-hidden="true"></i></a></div>
 +
                                            <div class="cr"><a ><i class="fa fa-circle" aria-hidden="true"></i></a></div>
 +
                                            <div class="cr"><a ><i class="fa fa-circle" aria-hidden="true"></i></a></div>
 +
                                            <div class="cr"><a ><i class="fa fa-circle" aria-hidden="true"></i></a></div>
 +
                                            <div class="cr"><a ><i class="fa fa-circle" aria-hidden="true"></i></a></div>
 +
                                        </div>                             
 +
                                    </div>     
 +
                                </div>
 +
                                <div class="col-xs-12 picture">
 
                                     <img src="https://static.igem.org/mediawiki/2018/4/40/T--Tianjin--measurement1.jpg">
 
                                     <img src="https://static.igem.org/mediawiki/2018/4/40/T--Tianjin--measurement1.jpg">
 
                                     <p>Figure 1. Mechanism of fluorescent proteins</p>
 
                                     <p>Figure 1. Mechanism of fluorescent proteins</p>
Line 201: Line 250:
 
                                     <p>
 
                                     <p>
 
                                       Luciferase is a popular choice as a reporter for gene expression because functional enzyme is created immediately upon translation and the assay is rapid, reliable and easy to perform.<sup><a href="#re1">[1,2]</a></sup> The fluorescent output of luciferase gene is due to the chemical reaction between luciferase and corresponding substrate, which can be easily detected by the luminoskan microplate reader.<br>
 
                                       Luciferase is a popular choice as a reporter for gene expression because functional enzyme is created immediately upon translation and the assay is rapid, reliable and easy to perform.<sup><a href="#re1">[1,2]</a></sup> The fluorescent output of luciferase gene is due to the chemical reaction between luciferase and corresponding substrate, which can be easily detected by the luminoskan microplate reader.<br>
                                       We used firefly luciferase(Fluc) gene purchased from Promega and Nanoluc luciferase(Nluc)  gene submitted by Team Tuebingen 2015 as another two reporters for our KaiABC system.
+
                                       We used firefly luciferase(Fluc) gene purchased from Promega and NanoLuc luciferase(Nluc)  gene submitted by Team Tuebingen 2015 as another two reporters for our KaiABC system.
 
                                     </p>
 
                                     </p>
 
                                 </div>
 
                                 </div>
Line 215: Line 264:
 
                                         </tbody>
 
                                         </tbody>
 
                                     </table>
 
                                     </table>
 +
                                </div>
 +
                                <div class="col-md-12 col-xs-12">
 +
                                    <div class="wrap">
 +
                                        <div class="photo-container">
 +
                                            <div class="display-left" onclick="" style="right:0px">
 +
                                                <a>&#10094;</a>
 +
                                            </div>
 +
                                            <div class="Slides">
 +
                                                <div class="mySlides">
 +
                                                    <img src="https://static.igem.org/mediawiki/2018/6/68/T--Tianjin--comic2.1.jpg">     
 +
                                                </div>
 +
                                                <div class="mySlides">
 +
                                                    <img src="https://static.igem.org/mediawiki/2018/3/37/T--Tianjin--comic2.2.jpg" >         
 +
                                                </div>
 +
                                                <div class="mySlides">
 +
                                                    <img src="https://static.igem.org/mediawiki/2018/5/5e/T--Tianjin--comic2.3.jpg" >         
 +
                                                </div>
 +
                                                <div class="mySlides">
 +
                                                    <img src="https://static.igem.org/mediawiki/2018/7/70/T--Tianjin--comic2.4.jpg" >         
 +
                                                </div>
 +
                                                <div class="mySlides">
 +
                                                    <img src="https://static.igem.org/mediawiki/2018/a/a7/T--Tianjin--comic2.5.jpg" >         
 +
                                                </div>             
 +
                                                <div class="mySlides">
 +
                                                    <img src="https://static.igem.org/mediawiki/2018/4/49/T--Tianjin--comic2.6.jpg" >
 +
                                                </div>
 +
                                                <div class="mySlides">
 +
                                                    <img src="https://static.igem.org/mediawiki/2018/0/04/T--Tianjin--comic2.7.jpg" >
 +
                                                </div>
 +
                                                <div class="mySlides">
 +
                                                    <img src="https://static.igem.org/mediawiki/2018/0/08/T--Tianjin--comic2.8.jpg" >
 +
                                                </div>
 +
                                                <div class="mySlides">
 +
                                                    <img src="https://static.igem.org/mediawiki/2018/9/94/T--Tianjin--comic2.9.jpg" >
 +
                                                </div>
 +
                                                <div class="mySlides">
 +
                                                    <img src="https://static.igem.org/mediawiki/2018/0/07/T--Tianjin--comic2.10.jpg" >
 +
                                                </div>
 +
                                            </div>
 +
                                            <div class="display-right" onclick="" style="left:0px;">
 +
                                                <a>&#10095;</a>
 +
                                            </div>
 +
                                        </div>
 +
                                        <div class="control">
 +
                                            <div class="cr"><a><i class="fa fa-circle" aria-hidden="true"></i></a></div>
 +
                                            <div class="cr"><a><i class="fa fa-circle" aria-hidden="true"></i></a></div>
 +
                                            <div class="cr"><a><i class="fa fa-circle" aria-hidden="true"></i></a></div>
 +
                                            <div class="cr"><a ><i class="fa fa-circle" aria-hidden="true"></i></a></div>
 +
                                            <div class="cr"><a ><i class="fa fa-circle" aria-hidden="true"></i></a></div>
 +
                                            <div class="cr"><a ><i class="fa fa-circle" aria-hidden="true"></i></a></div>
 +
                                            <div class="cr"><a ><i class="fa fa-circle" aria-hidden="true"></i></a></div>
 +
                                            <div class="cr"><a ><i class="fa fa-circle" aria-hidden="true"></i></a></div>
 +
                                            <div class="cr"><a ><i class="fa fa-circle" aria-hidden="true"></i></a></div>
 +
                                            <div class="cr"><a ><i class="fa fa-circle" aria-hidden="true"></i></a></div>
 +
                                        </div>                             
 +
                                    </div>     
 
                                 </div>
 
                                 </div>
 
                                 <div class="col-xs-12 picture">
 
                                 <div class="col-xs-12 picture">
Line 254: Line 359:
 
                                       When reconstructed in <em>Escherichia coli</em>,&nbsp;KaiABC&nbsp;oscillator showed a periodicity of 24 hours<sup><a href="#re3">[3]</a></sup>. To best verify our system can function in <em>Saccharomyces cerevisiae</em>&nbsp;, we planned to measure our system using different reporter at <strong>3-hour intervals for 1 day, 2 days, 3 days respectively</strong><strong>.</strong><br>
 
                                       When reconstructed in <em>Escherichia coli</em>,&nbsp;KaiABC&nbsp;oscillator showed a periodicity of 24 hours<sup><a href="#re3">[3]</a></sup>. To best verify our system can function in <em>Saccharomyces cerevisiae</em>&nbsp;, we planned to measure our system using different reporter at <strong>3-hour intervals for 1 day, 2 days, 3 days respectively</strong><strong>.</strong><br>
 
                                       As for the measurement of fluorescent proteins, we not only used the common detecting instrument, fourescence spectrophotometer, but also make an attempt to record the fluorescent variation via the cutting-edge Leica DMi8 inverted microscope.<br>
 
                                       As for the measurement of fluorescent proteins, we not only used the common detecting instrument, fourescence spectrophotometer, but also make an attempt to record the fluorescent variation via the cutting-edge Leica DMi8 inverted microscope.<br>
                                       Another thing worth mentioning is that, we bought all of the luciferase assay system from company Promega, among them there was a new kind of Nanoluc&nbsp;luciferase system----Nano-Glo<sup>&reg;</sup>&nbsp;Live Cell Assay System, which allows experimenters to detect live cell luminescent signals without lytic process, suitable for the 3-day assay. Despite there is no reference indicating that this product, a commonly used assay system in mammalian cells, could be used in<em>&nbsp;</em><em>Saccharomyces cerevisiae</em>, we decided to take a try and have successfully proved its validity. Consequently, we chose Nluc solely to complete our long time luminescent detection .
+
                                       Another thing worth mentioning is that, we bought all of the luciferase assay system from company Promega, among them there was a new kind of NanoLuc&nbsp;luciferase system----Nano-Glo<sup>&reg;</sup>&nbsp;Live Cell Assay System, which allows experimenters to detect live cell luminescent signals without lytic process, suitable for the 3-day assay. Despite there is no reference indicating that this product, a commonly used assay system in mammalian cells, could be used in<em>&nbsp;</em><em>Saccharomyces cerevisiae</em>, we decided to take a try and have successfully proved its validity. Consequently, we chose Nluc solely to complete our long time luminescent detection .
 
                                     </p>
 
                                     </p>
 
                                 </div>
 
                                 </div>
Line 265: Line 370:
 
                                         </thead>
 
                                         </thead>
 
                                         <tbody>
 
                                         <tbody>
                                             <tr><tr><td rowspan="4" width="154"><p><strong>Experimental</strong></p><br><br><p><strong>Group</strong></p></td><td width="468"><p>pABaC +pCiRbS +gal1+mCherry</p></td></tr><tr><td width="468"><p>pABaC +pbCiRS +gal1+mCherry</p></td></tr><tr><td width="468"><p>pABaC +pCiRbS+gal1+EYFP</p></td></tr><tr><td width="468"><p>pABaC +pbCiRS +gal1+EYFP</p></td></tr><tr><td rowspan="6" width="154"><p><strong>Negative</strong></p><br><br><p><strong>Control Group</strong></p></td><td width="468"><p>pABaC +gal1+mCherry</p></td></tr><tr><td width="468"><p>pCiRbS +gal1+mCherry</p></td></tr><tr><td width="468"><p>pbCiRS +gal1+mCherry</p></td></tr><tr><td width="468"><p>pABaC +gal1+EYFP</p></td></tr><tr><td width="468"><p>pbCiRS +gal1+EYFP</p></td></tr><tr><td width="468"><p>pCiRbS +gal1+EYFP</p></td></tr>
+
                                             <tr><tr><td rowspan="4" width="154"><p><strong>Experimental</strong></p><br><br><p><strong>Group</strong></p></td><td width="468"><p>pABaC +pCiRbS +p1m</p></td></tr><tr><td width="468"><p>pABaC +pbCiRS +p1m</p></td></tr><tr><td width="468"><p>pABaC +pCiRbS+p1E</p></td></tr><tr><td width="468"><p>pABaC +pbCiRS +p1E</p></td></tr><tr><td rowspan="6" width="154"><p><strong>Negative</strong></p><br><br><p><strong>Control Group</strong></p></td><td width="468"><p>pABaC +p1m</p></td></tr><tr><td width="468"><p>pCiRbS +p1m</p></td></tr><tr><td width="468"><p>pbCiRS +p1m</p></td></tr><tr><td width="468"><p>pABaC +p1E</p></td></tr><tr><td width="468"><p>pbCiRS +p1E</p></td></tr><tr><td width="468"><p>pCiRbS p1E</p></td></tr>
 
                                         </tbody>
 
                                         </tbody>
 
                                     </table>
 
                                     </table>
Line 281: Line 386:
 
                                         </thead>
 
                                         </thead>
 
                                         <tbody>
 
                                         <tbody>
                                             <tr><td width="154"><p><strong>Experimental</strong></p><p><strong>Group</strong></p></td><td width="468"><p>pABaC +pCiRbS +gal1+Fluc</p></td></tr><tr><td width="154"><p><strong>&nbsp;</strong></p></td><td width="468"><p>pABaC +pbCiRS +gal1+Fluc</p></td></tr><tr><td width="154"><p><strong>&nbsp;</strong></p></td><td width="468"><p>pABaC +pCiRbS +gal2+Nanoluc</p></td></tr><tr><td width="154"><p><strong>&nbsp;</strong></p></td><td width="468"><p>pABaC +pbCiRS +gal2+Nanoluc</p></td></tr><tr><td width="154"><p><strong>Negative</strong></p><p><strong>Control Group</strong></p></td><td width="468"><p>pABaC +gal1+Fluc</p></td></tr><tr><td width="154"><p><strong>&nbsp;</strong></p></td><td width="468"><p>pbCiRS +gal1+Fluc</p></td></tr><tr><td width="154"><p><strong>&nbsp;</strong></p></td><td width="468"><p>pCiRbS +gal1+Fluc</p></td></tr><tr><td width="154"><p><strong>&nbsp;</strong></p></td><td width="468"><p>pABaC +gal2+Nanoluc</p></td></tr><tr><td width="154"><p><strong>&nbsp;</strong></p></td><td width="468"><p>pbCiRS +gal2+Nanoluc</p></td></tr><tr><td width="154"><p><strong>&nbsp;</strong></p></td><td width="468"><p>pCiRbS +gal2+Nanoluc</p></td></tr>
+
                                             <tr><td width="154"><p><strong>Experimental</strong></p><p><strong>Group</strong></p></td><td width="468"><p>pABaC +pCiRbS +p1F</p></td></tr><tr><td width="154"><p><strong>&nbsp;</strong></p></td><td width="468"><p>pABaC +pbCiRS +p1F</p></td></tr><tr><td width="154"><p><strong>&nbsp;</strong></p></td><td width="468"><p>pABaC +pCiRbS +p2N</p></td></tr><tr><td width="154"><p><strong>&nbsp;</strong></p></td><td width="468"><p>pABaC +pbCiRS +p2N</p></td></tr><tr><td width="154"><p><strong>Negative</strong></p><p><strong>Control Group</strong></p></td><td width="468"><p>pABaC +p1F</p></td></tr><tr><td width="154"><p><strong>&nbsp;</strong></p></td><td width="468"><p>pbCiRS +p1F</p></td></tr><tr><td width="154"><p><strong>&nbsp;</strong></p></td><td width="468"><p>pCiRbS +p1F</p></td></tr><tr><td width="154"><p><strong>&nbsp;</strong></p></td><td width="468"><p>pABaC +p2N</p></td></tr><tr><td width="154"><p><strong>&nbsp;</strong></p></td><td width="468"><p>pbCiRS +p2N</p></td></tr><tr><td width="154"><p><strong>&nbsp;</strong></p></td><td width="468"><p>pCiRbS +p2N</p></td></tr>
 
                                         </tbody>
 
                                         </tbody>
 
                                     </table>
 
                                     </table>
Line 301: Line 406:
 
                                         </thead>
 
                                         </thead>
 
                                         <tbody>
 
                                         <tbody>
                                             <tr><td rowspan="4" width="154"><p><strong>Experimental</strong></p><p><strong>Group</strong></p></td><td width="468"><p>pABaC +pCiRbS +gal1+Nanoluc</p></td></tr><tr><td width="468"><p>pABaC +pbCiRS +gal1+Nanoluc</p></td></tr><tr><td width="468"><p>pABaC +pCiRbS +gal2+Nanoluc</p></td></tr><tr><td width="468"><p>pABaC +pbCiRS 1+gal2+Nanoluc</p></td></tr><tr><td rowspan="4" width="154"><p><strong>Control Group</strong></p></td><td width="468"><p>pABaC +gal2+Nanoluc</p></td></tr><tr><td width="468"><p>pbCiRS +gal2+Nanoluc</p></td></tr><tr><td width="468"><p>pCiRbS +gal2+Nanoluc</p></td></tr><tr><td width="468"><p>TDH3P+NanoLuc</p></td></tr>
+
                                             <tr><td rowspan="4" width="154"><p><strong>Experimental</strong></p><p><strong>Group</strong></p></td><td width="468"><p>pABaC +pCiRbS +p1N</p></td></tr><tr><td width="468"><p>pABaC +pbCiRS +p1N</p></td></tr><tr><td width="468"><p>pABaC +pCiRbS +p2N</p></td></tr><tr><td width="468"><p>pABaC +pbCiRS 1+p2N</p></td></tr><tr><td rowspan="4" width="154"><p><strong>Control Group</strong></p></td><td width="468"><p>pABaC +p2N</p></td></tr><tr><td width="468"><p>pbCiRS +p2N</p></td></tr><tr><td width="468"><p>pCiRbS +p2N</p></td></tr><tr><td width="468"><p>TDH3P+NanoLuc</p></td></tr>
 
                                         </tbody>
 
                                         </tbody>
 
                                     </table>
 
                                     </table>
Line 317: Line 422:
 
                                     </p>
 
                                     </p>
 
                                 </div>
 
                                 </div>
                                 <div class="col-xs-12 picture">
+
                                 <div class="col-xs-12 picture" id="M1">
 
                                     <img src="https://static.igem.org/mediawiki/2018/2/2e/T--Tianjin--EYFP123.jpg">
 
                                     <img src="https://static.igem.org/mediawiki/2018/2/2e/T--Tianjin--EYFP123.jpg">
                                     <p>Figure 6. pABaC +pCiRbS+gal1+EYFP photoed by normal microscope and Leica DMi8 </p>
+
                                     <p>Figure 6. pABaC +pCiRbS+p1E photoed by normal microscope and Leica DMi8 </p>
 
                                 </div>
 
                                 </div>
 
                             </div>
 
                             </div>
Line 328: Line 433:
 
         </div>
 
         </div>
  
        <div class="row">
+
                             
            <div class="panel-group" id="accordion3" role="tablist" aria-multiselectable="true">
+
                <div class="panel panel-default text-panel">
+
                    <div class="pan-heading text-pan-heading" id="collapsehead">
+
                        <div class="panel-title">
+
                            <a href="#collapseThree" role="button" data-toggle="collapse" data-parent="#accordion3" style="text-decoration: none;">
+
                                Measuement result
+
                            </a>
+
                        </div>
+
                    </div>
+
                    <div id="collapseThree" class="panel-collapse collapse" role="tabpanel" aria-labelledby="collapsehead" aria-expanded="false"">
+
                        <div class="panel-body">
+
                          <div class="row">
+
                                <div class="col-xs-12 picture">
+
                                    <img src="https://static.igem.org/mediawiki/2018/1/12/T--Tianjin--measurement7.jpg">
+
                                    <p>Figure 7. Experimental result reported by pABaC+pbCiRS+gal1+Fluc</p>
+
                                </div>
+
                                <div class="col-xs-12 picture">
+
                                    <img src="https://static.igem.org/mediawiki/2018/4/45/T--Tianjin--measurement8.jpg">
+
                                    <p>Figure 8. Experimental result reported by pABaC+pCiRbS+gal1+Fluc</p>
+
                                </div>
+
                                <div class="col-xs-12 picture">
+
                                    <img src="https://static.igem.org/mediawiki/2018/2/27/T--Tianjin--measurement9.jpg">
+
                                    <p>Figure 9. Experimental result reported by pABaC+pbCiRS+gal1+Nluc<br>(obtained by live cell assays)</p>
+
                                </div>
+
                                <div class="col-xs-12 picture">
+
                                    <img src="https://static.igem.org/mediawiki/2018/3/36/T--Tianjin--measurement10.jpg">
+
                                    <p>Figure 10.Experimental result reported by pABaC+pCiRbS+gal1+Nluc<br>(obtained by live cell assays)</p>
+
                                </div>
+
                                <div class="col-xs-12 picture">
+
                                    <img src="https://static.igem.org/mediawiki/2018/d/d6/T--Tianjin--measurement11.jpg">
+
                                    <p>Figure 11.Experimental result reported by pABaC+pbCiRS+gal2+Nluc<br>(obtained by live cell assays)</p>
+
                                </div>
+
                                <div class="col-xs-12 picture">
+
                                    <img src="https://static.igem.org/mediawiki/2018/7/7a/T--Tianjin--measurement12.jpg">
+
                                    <p>Figure 12.Experimental result reported by pABaC+pCiRbS+gal2+Nluc<br>(obtained by live cell assays)</p>
+
                                </div>
+
                                <div class="col-xs-12 text">
+
                                    <p>Some basic information can be gained from the above figures:</p>
+
                                    <ol>
+
                                        <li>Compared with the corresponding control groups, all the experimental groups have showed apparent oscillatory signals.</li>
+
                                        <li>In the measured 3500 min, the oscillations in 700-2000 min are extremely unstable and nearly absent, while the oscillations for the rest of the time are relatively stable.</li>
+
                                        <li>The cycle is about 500min (about 8.3h).</li>
+
                                    </ol>
+
                                    <p><strong><strong>Note: final measurement results and analysis should be accessed in <a href="https://2018.igem.org/Team:Tianjin/Demonstrate">Demonstrate</a> page.</strong></strong></p>
+
                                </div>
+
 
                             </div>
 
                             </div>
 
                         </div>
 
                         </div>
Line 463: Line 523:
 
     </div>
 
     </div>
 
<script type="text/javascript" src="https://2018.igem.org/Team:Tianjin/js/timeline?action=raw&ctype=text/javascript"></script>
 
<script type="text/javascript" src="https://2018.igem.org/Team:Tianjin/js/timeline?action=raw&ctype=text/javascript"></script>
 +
<script src="https://2018.igem.org/Team:Tianjin/js/slides?action=raw&ctype=text/javascript"></script>
 
</body>
 
</body>
 
</html>
 
</html>

Latest revision as of 13:47, 13 December 2018

<!DOCTYPE html> Team:Tianjin - 2018.igem.org

MEASUREMENT

In pursuit of a rigorously scientific result, we have adopted multiple measuring approaches throughout the whole project, among which two predominant methods described in extreme detail in wiki, would inspire relative researchers.

As two sophisticated and renowned reporters in biotechnological domain, fluorescent protein and luciferase were opted to illustrate the effectiveness of our constructed oscillating system, detected by combined means and instrument. Mcherry and EYFP, two selected fluorescent reporters were measured by both fluorescence spectrophotometer and Leica DMi8 respectively. Leica DMi8 allows us to visually and intuitively see the expression of the fluorescent protein EYFP in Saccharomyces cerevisiae In terms of luciferase, in addition to commonly used firefly luciferase, a courageous attempt we made was to choose a novel luciferase NanoLuc and its live cell substrate in spite of no records of the usefulness in Saccharomyces cerevisiae before and obtained expected outcome successfully.