Difference between revisions of "Team:ZJU-China/Human Practices"

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                           <a href="#" class="dropdown-toggle link" data-toggle="dropdown">Overview<b class="caret"></b></a>
 
                           <a href="#" class="dropdown-toggle link" data-toggle="dropdown">Overview<b class="caret"></b></a>
 
                           <ul class="dropdown-menu ">
 
                           <ul class="dropdown-menu ">
−
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Overview">Project Description</a></li>
+
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Overview">Description</a></li>
 +
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Demonstrate">Demonstrate</a></li>
 +
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Applied_Design">Applied Design</a></li>
 
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Achievements">Achievements</a></li>
 
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Achievements">Achievements</a></li>
 +
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Improve">Improve Parts</a></li>
 
                               <li><a href="https://2017.igem.org/Team:ZJU-China/InterLab">InterLab</a></li>
 
                               <li><a href="https://2017.igem.org/Team:ZJU-China/InterLab">InterLab</a></li>
−
                              <li><a href="https://2017.igem.org/Team:ZJU-China/ImproveParts">Improve Parts</a></li>
+
 
 
                           </ul>
 
                           </ul>
 
                       </li>
 
                       </li>
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                           <a href="#" class="dropdown-toggle link" data-toggle="dropdown">Project<b class="caret"></b></a>
 
                           <a href="#" class="dropdown-toggle link" data-toggle="dropdown">Project<b class="caret"></b></a>
 
                           <ul class="dropdown-menu ">
 
                           <ul class="dropdown-menu ">
−
                              <!--<li><a href="https://2017.igem.org/Team:ZJU-China/Project">Project Home</a></li>-->
 
 
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Project/tp">Trichoderma Proof</a></li>
 
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Project/tp">Trichoderma Proof</a></li>
 
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Project/voc">VOC sensors</a></li>
 
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Project/voc">VOC sensors</a></li>
−
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Project/st">Signal Transduction</a></li>
+
                               <li><a style="font-size: 0.7em!important;" href="https://2017.igem.org/Team:ZJU-China/Project/st">Chemical Signal Transduction</a></li>
−
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Project/ms">Mat Synthesis</a></li>
+
                               <li><a style="font-size: 0.7em!important;" href="https://2017.igem.org/Team:ZJU-China/Project/mt">Medium Wave Transduction</a></li>
−
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Project/conclusion">Conclusion</a></li>
+
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Project/Downstream">Downstream</a></li>
−
                               <!--<li><a href="https://2017.igem.org/Team:ZJU-China/Project/improvement">Improvement</a></li>-->
+
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Project/conclusion">Conclusions</a></li>
−
                               <!--<li><a href="https://2017.igem.org/Team:ZJU-China/InterLab">Interlab</a></li>-->
+
 
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Notebook">Notebook</a></li>
 
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Notebook">Notebook</a></li>
 +
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Protocols">Protocols</a></li>
 
                           </ul>
 
                           </ul>
 
                       </li>
 
                       </li>
  
 
                       <li class="m_nav_item dropdown" >
 
                       <li class="m_nav_item dropdown" >
−
                           <a href="#" class="dropdown-toggle link" data-toggle="dropdown">Modelling<b class="caret"></b></a>
+
                           <a href="#" class="dropdown-toggle link" data-toggle="dropdown">Model<b class="caret"></b></a>
 
                           <ul class="dropdown-menu ">
 
                           <ul class="dropdown-menu ">
 
                               <!--<li><a href="https://2017.igem.org/Team:ZJU-China/Model">Summery</a></li>-->
 
                               <!--<li><a href="https://2017.igem.org/Team:ZJU-China/Model">Summery</a></li>-->
−
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Model">Coculture</a></li>
+
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Model">VOC analysis</a></li>
−
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Model/VOC">VOC analysis</a></li>
+
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Model/Coculture">Coculture</a></li>
  
 
                           </ul>
 
                           </ul>
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                           <ul class="dropdown-menu ">
 
                           <ul class="dropdown-menu ">
 
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Parts">All Parts</a></li>
 
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Parts">All Parts</a></li>
−
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Parts/Basic">Basic Parts</a></li>
+
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Basic_Part">Basic Parts</a></li>
−
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Parts/Composite">Composite Parts</a></li>
+
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Composite_Part">Composite Parts</a></li>
−
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Parts/Collection">Parts Collection</a></li>
+
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Part_Collection">Part Collection</a></li>
 +
 
 +
 
 +
 
 +
 
 
                           </ul>
 
                           </ul>
 
                       </li>
 
                       </li>
  
−
                       <li><a href="https://2017.igem.org/Team:ZJU-China/Hardware">Hardware</a></li>
+
                       <li class="m_nav_item dropdown" >
 +
                          <a href="#" class="dropdown-toggle link" data-toggle="dropdown">Hardware<b class="caret"></b></a>
 +
                          <ul class="dropdown-menu ">
 +
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Hardware">Overview</a></li>
 +
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Hardware/Device">Device</a></li>
 +
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Hardware/Improvements">Improvements</a></li>
 +
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Hardware/MediumWave">Medium Wave</a></li>
 +
                          </ul>
 +
                      </li>
  
 
                       <li class="m_nav_item dropdown" >
 
                       <li class="m_nav_item dropdown" >
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                           </ul>
 
                           </ul>
 
                       </li>
 
                       </li>
−
 
−
                      <li><a href="https://2017.igem.org/Team:ZJU-China/HP/Collaborations">Collaborations</a></li>
 
  
 
                       <li class="m_nav_item dropdown">
 
                       <li class="m_nav_item dropdown">
 
                           <a href="#" class="dropdown-toggle link" data-toggle="dropdown">HP<b class="caret"></b></a>
 
                           <a href="#" class="dropdown-toggle link" data-toggle="dropdown">HP<b class="caret"></b></a>
 
                           <ul class="dropdown-menu ">
 
                           <ul class="dropdown-menu ">
 +
 
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Human_Practices">Summary</a></li>
 
                               <li><a href="https://2017.igem.org/Team:ZJU-China/Human_Practices">Summary</a></li>
 
                               <li><a href="https://2017.igem.org/Team:ZJU-China/HP/Silver">Silver</a></li>
 
                               <li><a href="https://2017.igem.org/Team:ZJU-China/HP/Silver">Silver</a></li>
−
                               <li><a href="https://2017.igem.org/Team:ZJU-China/HP/Gold_Integrated">Gold</a></li>
+
                               <li><a href="https://2017.igem.org/Team:ZJU-China/HP/Gold_Integrated">Gold Integrated</a></li>
 +
                              <li><a href="https://2017.igem.org/Team:ZJU-China/Engagement">Engagement</a></li>
 
                           </ul>
 
                           </ul>
 
                       </li>
 
                       </li>
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                           </ul>
 
                           </ul>
 
                       </li>
 
                       </li>
−
 
 
                   </ul>
 
                   </ul>
 
               </div>
 
               </div>
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<div style="margin:0 auto;z-index:50;">
 
<div style="margin:0 auto;z-index:50;">
−
   <img src="https://static.igem.org/mediawiki/2017/b/be/ZJUChina_interlab_banner.jpg" width="100%">
+
   <img src="https://static.igem.org/mediawiki/2017/1/1b/ZJU_China_HPPublic_248.jpg" width="100%">
 
   <!-- <img src="images/HP_temp/banner.jpg" width="100%"> -->
 
   <!-- <img src="images/HP_temp/banner.jpg" width="100%"> -->
 
</div>
 
</div>
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<div class="timelineLight tl">
 
<div class="timelineLight tl">
 
     <div class="item" data-id="23/05/2017" data-description="Software">
 
     <div class="item" data-id="23/05/2017" data-description="Software">
−
       <img class="con_borderImage" src="images/hp/timeline/1.jpg" alt="" />
+
       <img class="con_borderImage" src="https://static.igem.org/mediawiki/2017/3/3c/ZJU_China_HP_wx.png" alt="" />
 
       <h2 class="timelineHead">MAY, 23</h2>
 
       <h2 class="timelineHead">MAY, 23</h2>
 
       <span>When your team is on the stage of brainstorming, are you bothered with looking for the former iGEM teams' information? Take out your smart phone and try our Mini Program.
 
       <span>When your team is on the stage of brainstorming, are you bothered with looking for the former iGEM teams' information? Take out your smart phone and try our Mini Program.
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       <img class="con_borderImage" src="https://static.igem.org/mediawiki/2017/3/33/ZJU_China_HP_high.jpg" alt="" />
 
       <img class="con_borderImage" src="https://static.igem.org/mediawiki/2017/3/33/ZJU_China_HP_high.jpg" alt="" />
 
       <h2 class="timelineHead">MAY, 28</h2>
 
       <h2 class="timelineHead">MAY, 28</h2>
−
       <span>"We just built up a iGEM high school team, how shall we start?"
+
       <span>"We just built up a iGEM high school team, how shall we start?"<br>
 +
"Don't worry! We can share our experience with you."
 +
 
  
 
</span>
 
</span>
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−
     <div class="item" data-id="04/07/2017" data-description="Lorem ipsum dolor sit amet, consectetur adipiscing elit...">
+
     <div class="item" data-id="04/07/2017" data-description="Design">
−
       <img class="con_borderImage" src="images/hp/timeline/3.jpg" alt="" />
+
       <img class="con_borderImage" src="https://static.igem.org/mediawiki/2017/c/cf/ZJU_China_HP_zbsmall.jpg" alt="" />
 
       <h2 class="timelineHead">JULY, 4</h2>
 
       <h2 class="timelineHead">JULY, 4</h2>
−
       <span>ZHOU BIAN...</span>
+
       <span>After determining our logo, we designed some derivative products such as postcard, key chain, badge, notebook, stickers, etc. We sincerely hope that these gifts can get everyone's favor during intercollegiate communication.</span>
 
     </div>
 
     </div>
  
  
 
     <div class="item" data-id="09/07/2017" data-description="The hall in ZJSTM">
 
     <div class="item" data-id="09/07/2017" data-description="The hall in ZJSTM">
−
       <img class="con_borderImage" src="https://static.igem.org/mediawiki/2017/f/f6/ZJU_China_HP_kxy.jpg" alt="" />
+
       <img class="con_borderImage" src="https://static.igem.org/mediawiki/2017/2/28/ZJU_China_HP_kjg.jpg" alt="" />
 
       <h2 class="timelineHead">JULY, 9</h2>
 
       <h2 class="timelineHead">JULY, 9</h2>
−
       <span>We held a "garden party" in the Zhejiang Science and Technology Museum, the children who joined in it said: "I've learned lots of scientific knowledge in this activity." "I found it interesting. Give the thumbs up."</span>
+
       <span>We held a "garden party" in the Zhejiang Science and Technology Museum. The children who joined in it said: "I've learned lots of scientific knowledge in this activity." "I found it interesting. Give the thumbs up."</span>
 
     </div>
 
     </div>
 
      
 
      
 
      
 
      
−
     <div class="item" data-id="13/07/2017" data-description="yao jian ju">
+
     <div class="item" data-id="13/07/2017" data-description="State Food and Drug Administration">
 
       <img class="con_borderImage" src="https://static.igem.org/mediawiki/2017/1/13/ZJU_China_HP_yjj.jpg" alt="" />
 
       <img class="con_borderImage" src="https://static.igem.org/mediawiki/2017/1/13/ZJU_China_HP_yjj.jpg" alt="" />
 
       <h2 class="timelineHead">JULY, 13</h2>
 
       <h2 class="timelineHead">JULY, 13</h2>
−
       <span>yaojianju</span>
+
       <span>"I do believe that such cutting edge technology can do great benefits to people once it grows fully mature. But we have to make sure it's safe before we take a closer look at its effectiveness," one of the officials said.</span>
 
     </div>
 
     </div>
 
      
 
      
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     <div class="item" data-id="19/07/2017" data-description="National Demonstration Center for Experimental Biology Education in ZJU">
 
     <div class="item" data-id="19/07/2017" data-description="National Demonstration Center for Experimental Biology Education in ZJU">
−
       <img class="con_borderImage" src="https://static.igem.org/mediawiki/2017/2/28/ZJU_China_HP_kjg.jpg" alt="" />
+
       <img class="con_borderImage" src="https://static.igem.org/mediawiki/2017/f/f6/ZJU_China_HP_kxy.jpg" alt="" />
 
       <h2 class="timelineHead">JULY, 19</h2>
 
       <h2 class="timelineHead">JULY, 19</h2>
−
       <span>"Wow, What is this? How shall I use it?"
+
       <span>National Demonstration Center for Experimental Biology Education in ZJU<br>"Wow, What is this? How shall I use it?"<br>"Welcome to our lab, let me guide you!"</span>
−
"Welcome to our lab, let me guide you!"
+
−
</span>
+
 
     </div>
 
     </div>
 
      
 
      
 
     <div class="item" data-id="20/07/2017" data-description="Software">
 
     <div class="item" data-id="20/07/2017" data-description="Software">
−
       <img class="con_borderImage" src="images/hp/timeline/7.jpg" alt="" />
+
       <img class="con_borderImage" src="https://static.igem.org/mediawiki/2017/1/13/ZJU_China_HP_Game.png" alt="" />
 
       <h2 class="timelineHead">JULY, 20</h2>
 
       <h2 class="timelineHead">JULY, 20</h2>
−
       <span>Can Trichoderma defend your plant from dangerous? It depends on you.  
+
       <span>Can Trichoderma defend your plant from dangerous? It depends on you. Play our computer game and see who's the winner.</span>
−
Play our computer game and see who's the winner.
+
−
</span>
+
 
     </div>
 
     </div>
 
      
 
      
−
     <div class="item" data-id="23/08/2017" data-description="yan cao gong si">
+
     <div class="item" data-id="23/08/2017" data-description="Hunan Province">
 
       <img class="con_borderImage" src="https://static.igem.org/mediawiki/2017/5/59/ZJU_China_HP_ycgs.jpg" alt="" />
 
       <img class="con_borderImage" src="https://static.igem.org/mediawiki/2017/5/59/ZJU_China_HP_ycgs.jpg" alt="" />
 
       <h2 class="timelineHead">AUGUST, 23</h2>
 
       <h2 class="timelineHead">AUGUST, 23</h2>
−
       <span>yan cao gong si</span>
+
       <span>By visiting the tobacco field as well as communicating with the officials in the tobacco company, we got useful suggestions and inspirations. Then we designed our device and modified our "wetlab" work.</span>
 +
    </div>
 +
 
 +
    <div class="item" data-id="25/08/2017" data-description="Northeastern University">
 +
        <img class="con_borderImage" src="https://static.igem.org/mediawiki/2017/5/5d/ZJU_China_HP_DBDX.jpg" alt="" />
 +
        <h2 class="timelineHead">AUGUST, 25</h2>
 +
        <span>We had a freewheeling talk with the iGEM team of Northeastern University, because of this talk,  we later had opportunities to collaborate with them in experiments.</span>
 
     </div>
 
     </div>
 
      
 
      
 
     <div class="item" data-id="26/08/2017" data-description="CCiC">
 
     <div class="item" data-id="26/08/2017" data-description="CCiC">
−
       <img class="con_borderImage" src="images/hp/timeline/9.jpg" alt="" />
+
       <img class="con_borderImage" src="https://static.igem.org/mediawiki/2017/5/52/ZJU_China_HP_CCIC.jpg" alt="" />
 
       <h2 class="timelineHead">AUGHST, 26</h2>
 
       <h2 class="timelineHead">AUGHST, 26</h2>
−
       <span>CCiC</span>
+
       <span>Many suggestions that we received from other teams are of value, so we further improved our design after coming back. Besides, we built close cooperation with UESTC-China since we both used the tobacco.</span>
 +
    </div>
 +
 
 +
    <div class="item" data-id="07/10/2017" data-description="BTGEM Summit">
 +
        <img class="con_borderImage" src="https://static.igem.org/mediawiki/2017/d/db/ZJU_China_HP_BTGEM.jpg" alt="" />
 +
        <h2 class="timelineHead">OCTOBER, 7</h2>
 +
        <span>We trained ourselves through telling others our story, and they fed back what they were interested in or what they could not figure out, and all these information helped us discover many defects in our presentation.</span>
 
     </div>
 
     </div>
 
      
 
      
Line 550: Line 551:
  
  
−
<div style="background-color: #FFFAF0;width: 100%" class="container zjuContent">
+
<div style="width: 100%" class="container zjuContent">
 
   <div class="col-md-3"></div>
 
   <div class="col-md-3"></div>
 
     <div class="col-md-9" role="main">
 
     <div class="col-md-9" role="main">
Line 556: Line 557:
  
 
    
 
    
−
   <h1 id="introduction" class="page-header ArticleHead GreenAH">Introduction</h1>
+
   <h1 id="summary" class="page-header ArticleHead GreenAH">Human Practice Summary</h1>
−
  <p class="PP">The InterLab study is developed by the Measurement Committee. Since synthetic biology, also been called engineering biology, is based on engineering thinking and methods, it values the reliability and repeatability of measurements. The InterLab study, involving teams and labs around the world，takes green fluorescent protein (gfp) as an example after one strict, detailed protocol to ensure the common, comparable units for measuring, which hopes to test the robustness by comparing the results from different labs.</p>
+
          <p class="PP">We engaged with stakeholders as well as experts,and did field study etc., which as a whole helped us to design and improve our project.</p>
−
  <p class="PP">This is the Fourth Measurement InterLab and our team participates in it. The InterLab this year tries to establish a GFP measurement protocol based on engineering principles, find out the similarity between fluorescence measured around the world and make gene expression more precise and reliable through tests of some RBS devices (BCDs). We are provided with materials for standard curves together with positive control, negative control and devices with different promoters and RBSs. Strictly following the requirements and protocol, we have obtained the relevant datas.</p>
+
          <p class="PP">We consider the safety problem and interviewed the officials about bio-safety legislation. New designs and changes occurred after we made a trip to the tobacco company. Also, we made a lot of connection with public, especially teenagers, through activities like garden party, open house day, project-related computer game and so on. What's more, we joined in some meetings with other participants.</p>
 +
          <p class="PP">We saw the current condition and problem, and made dialogues with professors and common people to got useful advice. Then, we made new things(like our VOC device) and confirmed one direction of our project. Through the project process, human practice helped us throw light on public issues and make efforts to solve the practical problem.</p>
  
−
<h1 id="method" class="page-header ArticleHead GreenAH">METHODS & MATERIALS</h1>
+
          <div style="text-align: center;">
−
<h2 id="transformation" class="H2Head">Transformation</h2>
+
              <br><br>
−
<p class="PP">Transform Escherichia coli DH5α with these following plasmids:</p>
+
          <a class="CuteButton GreenCB" href="https://2017.igem.org/Team:ZJU-China/HP/Silver">More HP About Silver...</a>
−
<p class="PP">
+
          <br><br><br>
−
<ul class="Retract">
+
          <a class="CuteButton YellowCB" href="https://2017.igem.org/Team:ZJU-China/HP/Gold_Integrated">More HP About Gold...</a>
−
<li>Positive control</li>
+
          </div>
−
<li>Negative control</li>
+
      </div>
−
<li>Test Device 1: J23101+I13504</li>
+
−
<li>Test Device 2: J23106+I13504</li>
+
−
<li>Test Device 3: J23117+I13504</li>
+
−
<li>Test Device 4: J23101.BCD2.E0040.B0015</li>
+
−
<li>Test Device 5: J23106.BCD2.E0040.B0015</li>
+
−
<li>Test Device 6: J23117.BCD2.E0040.B0015</li>
+
−
</ul></p>
+
−
<p class="PP">Resuspend DNA in selected wells in the Distribution Kit with 10µL ddH20. Thaw competent cells on ice. Pipette 25µL of competent cells into 1.5mL tube per transformation and add 2µL of resuspended DNA into it. Incubate on ice for 30min. Heat shock tubes at 42°C for 90 sec. Then incubate on ice for 5min.</p>
+
−
<p class="PP">Add 200µL SOC media with Chloramphenicol(1000×) to each transformation. Incubate at 37°C for 1 hours, shaking at 200-300rpm.</p>
+
−
<p class="PP">Pipette 100µL of each transformation onto LB plates(Chloramphenicol, 1000×). Spread with sterilized spreader. Incubate transformations overnight (14-18hr) at 37°C.</p>
+
−
 
+
−
<h2 id="colonies" class="H2Head">Colonies Selection</h2>
+
−
<p class="PP">Pick 2 single colonies from each of plate and inoculate it on 5-10 mL LB medium with Chloramphenicol(1000×). Grow the cells overnight (16-18 hours) at 37°C and 220 rpm. Check them under the fluorescence microscope before further experiments.</p>
+
−
 
+
−
<h2 id="calibration" class="H2Head">Calibration</h2>
+
−
<p class="PP">We used the plate reader Synergy Neo2 for all the measurements and we used black 96 well plates with flat, transparent bottom.</p>
+
−
 
+
−
<h3 class="H3Head">OD600 Reference Point</h3>
+
−
<p class="PP">Add 100 µl LUDOX into wells A1, B1, C1, D1 and 100 µl of H2O into wells A2, B2, C2, D2. Measure absorbance 600nm of all samples in all standard measurement modes in instrument, pathlength correction was turned off. The temperature setting was 24℃. Record the data.</p>
+
−
 
+
−
<h3 class="H3Head">Fluorescein Fluorescence Standard Curve</h3>
+
−
<p class="PP">Spin down fluorescein stock tube. Prepare 2x fluorescein stock solution (100 µM) by resuspending fluorescein in 1 mL of 1xPBS. Dilute the 2x fluorescein stock solution with 1xPBS to make a 1x fluorescein solution and resulting concentration of fluorescein stock solution 50 µM.</p>
+
−
<p class="PP">Prepare the serial dilutions of fluorescein as shown below. Set 4 copies.</p>
+
−
+
−
<img class="textimg" src="https://static.igem.org/mediawiki/2017/7/71/ZJUChina_interlab_screenshot1.png">
+
−
<p class="PP">Measure the plate in plate reader, the excitation filter was set to 485nm/5nm and the emission filter was set to 530nm/30nm. Pathlength correction was turned off. The gain setting was 70. Fluorescence was from the top. The temperature setting was 24℃. Record the data.</p>
+
−
 
+
−
<h2 id="cellmeasurement" class="H2Head">Cell Measurement</h2>
+
−
<p class="PP">Measure OD<sub>600</sub> of the overnight cultures prepared after colony selection, then dilute the culture to a target OD600 of 0.02 in LB medium + Chloramphenicol with the help of Excel(Dilution Calculation) sheet. Incubate the cultures at 37°C and 220 rpm. Take 500 µL samples of the cultures from each of the 8 devices, two colonies per device, at 0, 2, 4, and 6 hours of incubation and add them into 96 well plates as shown below. Place samples on ice before measurements</p>
+
−
<img class="textimg" src="https://static.igem.org/mediawiki/2017/a/a1/ZJUChina_interlab_screenshot2.png">
+
−
<p class="PP">Measure the samples (OD and Fl measurement). The cell measurement was under the same condition with fluorescein fluorescence standard curve and used the same plate.</p>
+
−
<h1 id="results" class="page-header ArticleHead GreenAH">Results</h1>
+
−
 
+
−
<h2 id="OD600" class="H2Head">OD600 Reference point</h2>
+
−
 
+
−
<div class="panel panel-default">
+
−
  <!-- Default panel contents -->
+
−
  <div class="panel-heading">Panel heading</div>
+
−
 
+
−
  <!-- Table -->
+
−
  <table class="table">
+
−
  <tr>
+
−
<th> </th>
+
−
<th>LUDOX-HS40</th>
+
−
<th>H<sub>2</sub>O</th>
+
−
</tr>
+
−
 
+
−
<tr>
+
−
<th>Replicate 1</th>
+
−
<td class="blueTable">0.05</td>
+
−
<td class="blueTable">0.035</td>
+
−
</tr>
+
−
 
+
−
<tr>
+
−
<th>Replicate 2</th>
+
−
<td class="blueTable">0.049</td>
+
−
<td class="blueTable">0.037</td>
+
−
</tr>
+
−
 
+
−
<tr>
+
−
<th>Replicate 3</th>
+
−
<td class="blueTable">0.049</td>
+
−
<td class="blueTable">0.039</td>
+
−
</tr>
+
−
 
+
−
<tr>
+
−
<th>Replicate 4</th>
+
−
<td class="blueTable">0.049</td>
+
−
<td class="blueTable">0.039</td>
+
−
</tr>
+
−
 
+
−
<tr>
+
−
<th>Arith. Mean</th>
+
−
<td class="yellowTable">0.04925</td>
+
−
<td class="yellowTable">0.0375</td>
+
−
</tr>
+
−
 
+
−
<tr>
+
−
<th>Corrected Abs600</th>
+
−
<td class="yellowTable">0.01175</td>
+
−
<td> </td>
+
−
</tr>
+
−
 
+
−
<tr>
+
−
<th>Reference OD600</th>
+
−
<td class="yellowTable">0.0425</td>
+
−
<td> </td>
+
−
</tr>
+
−
 
+
−
<tr>
+
−
<th>OD600/Abs600</th>
+
−
<td class="yellowTable">3.617021277</td>
+
−
<td> </td>
+
−
</tr>
+
−
 
+
−
  </table>
+
−
</div>
+
−
 
+
−
<h2 id="fluorescein" class="H2Head">Fluorescein Fluorescence Standard Curve</h2>
+
−
<img class="textimg" src="https://static.igem.org/mediawiki/2017/9/9c/ZJUChina_interlab_diagram1.png">
+
−
<img class="textimg" src="https://static.igem.org/mediawiki/2017/b/b2/ZJUChina_interlab_diagram2.png">
+
−
<h2 id="discussion" class="H2Head">Discussion</h2>
+
−
<img class="textimg" src="https://static.igem.org/mediawiki/2017/8/83/ZJUChina_interlab_diagram3.png">
+
−
<p class="PP">Abs600 of the samples changed over time as shown, in which the data of each sample is the average of 4 replicates. Abs600 of the samples reflected its concentration, which could show the growth of bacteria. As the picture shown, the growth rate of bacteria generally got slower as time went by, this is probably because at the beginning of the measurement, bacteria was in the later period of logarithmic phase and the resources were not that sufficient, inhibiting the growth. Several samples appeared to be different, such as colony 1 of device 4, which seemed to be in the logarithmic phase and colony 2 of device 1, whose concentration went down after 2 hours of incubation, perhaps low activity resulting in massive death.</p>
+
−
<p class="PP">In addition, growth curves of each colony were different. This may be caused by different initial concentration of samples after dilution, which led the samples be in different growing statuses. We will mention it later.</p>
+
−
<img class="textimg" src="https://static.igem.org/mediawiki/2017/6/6e/ZJUChina_interlab_diagram4.png">
+
−
<img class="textimg" src="https://static.igem.org/mediawiki/2017/3/3e/ZJUChina_interlab_diagram5.png">
+
−
<p class="PP">The fluorescence differed obviously. The absolute fluorescence and relative fluorescence of device 3, device 5 and device 6 were relatively low, for which their promoters and RBSs might account. For two replicates of device 1, their fluorescence differed greatly and the same as growth, making it hard to compare. Either operational error or colony selection would affect.</p><p class="PP">Furthermore, the tendence of absolute fluorescence turned out to rise and fall, mostly going down as time went by. We thought this was because the protein expression varied in different growth phase and growth of bacteria changed over time.</p>
+
−
<h2 id="reflection" class="H2Head">Reflection</h2>
+
−
<p class="PP">Some of our operations might affect the results, which can be improved.</p>
+
−
<p class="PP">Our lab do not have plate reader, so we borrowed one from other lab. We cultured bacteria and loaded samples in our lab and then took them to the other lab for measurement. Since it took some time to go back and forth, our actual measurement took longer than expectation, so the measurement postponed a little. Though samples were laid on ice, the bacteria could still growth, making the real measurement time different from the demand. Similarly, the cultures settled in the process, leading to the deviation of the concentration after dilution. As a result, the comparability between each sample was affected.</p>
+
−
<p class="PP">By the way, the first time we did the OD600 Reference Point measurement, we put the 96 well plate containing LUDOX on ice. Then it turned out that the OD600 of LUDOX was lower than water. Maybe LUDOX denatured in the cold environment on ice referred to the official warning. So perhaps it is better not to put LUDOX on ice or anywhere like that.</p>
+
−
<h1 id="feedback" class="page-header ArticleHead GreenAH">Feedback</h1>
+
−
<p class="PP">After the InterLab Study measurement, we have something to say about the provided protocol, which goes on like this:</p>
+
−
<h1 id="advantages" class="page-header ArticleHead YellowAH">Advantages</h1>
+
−
 
+
−
<ul id="improvements" class="Retract contentLi">
+
−
<li>The protocol is not only detailed but also clear, so it is easy to operate and avoid differences between teams.</li>
+
−
<li>The provided Excel sheets make it easier to calculate.</li>
+
−
<li>There are extra materials for mistakes and failure.</li>
+
−
<li>Some warnings are necessary, such as LUDOX cannot be stored in refrigerator and oversaturated detector will cause low concentrations linear, but high concentrations saturate or fall for Fluorescein standard curve. It helps a lot in finding solutions to problems.</li>
+
−
 
+
−
</ul>
+
−
 
+
−
 
+
−
<h1 class="page-header ArticleHead YellowAH">Improvements</h1>
+
−
<ul class="Retract contentLi">
+
−
<li>The PDF version of protocol differs from the Excel sheets in some ways, such as the volume of diluted culture. Though it won’t affect the operation and results, it can cause misunderstanding and confusion.</li>
+
−
<li>Excel sheets can list out the computational process, which hopes to contribute to data verification and data analysis.</li>
+
−
<li>More samples and replicates can be involved to avoid mistakes and the time span between measurements can be shortened.</li>
+
−
</ul>
+
−
<p class="PP">We sincerely hope the InterLab Study this year will success and hope the InterLab Study next year will go well.</p>
+
−
</div>
+
 
          
 
          
  
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−
       <li><a href="#introduction">Introduction</a></li>
+
       <li><a href="#summary">Summary</a></li>
−
      <li>
+
 
−
        <a href="#methods">Methods&Materials</a>
+
−
          <ul class="nav">
+
−
            <li><a href="#transformation">Transformation</a></li>
+
−
            <li><a href="#colonies">Colonies Selection</a></li>
+
−
            <li><a href="#calibration">Calibration</a></li>
+
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            <li><a href="#cellmeasurement">Cell Measurement</a></li>
+
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+
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+
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      <li>
+
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        <a href="#results">Results</a>
+
−
          <ul class="nav">
+
−
            <li><a href="#OD600">OD600</a></li>
+
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            <li><a href="#fluorescein">Fluorescein</a></li>
+
−
            <li><a href="#discussion">Discussion</a></li>
+
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+
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+
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+
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+
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+
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          <li><a href="#advantages">Advantages</a></li>
+
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          <li><a href="#improvements">Improvements</a></li>
+
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+
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+
 
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     </ul>
  
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+
      <a class="back-to-top" href="#top">
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  <!--  <i class="fa fa-arrow-up nav_icon"></i> -->
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          <!--  <i class="fa fa-arrow-up nav_icon"></i> -->
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    ↑ UP ↑
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Latest revision as of 02:09, 2 November 2017

HP

MAY, 23

When your team is on the stage of brainstorming, are you bothered with looking for the former iGEM teams' information? Take out your smart phone and try our Mini Program.

MAY, 28

"We just built up a iGEM high school team, how shall we start?"
"Don't worry! We can share our experience with you."

JULY, 4

After determining our logo, we designed some derivative products such as postcard, key chain, badge, notebook, stickers, etc. We sincerely hope that these gifts can get everyone's favor during intercollegiate communication.

JULY, 9

We held a "garden party" in the Zhejiang Science and Technology Museum. The children who joined in it said: "I've learned lots of scientific knowledge in this activity." "I found it interesting. Give the thumbs up."

JULY, 13

"I do believe that such cutting edge technology can do great benefits to people once it grows fully mature. But we have to make sure it's safe before we take a closer look at its effectiveness," one of the officials said.

JULY, 19

National Demonstration Center for Experimental Biology Education in ZJU
"Wow, What is this? How shall I use it?"
"Welcome to our lab, let me guide you!"

JULY, 20

Can Trichoderma defend your plant from dangerous? It depends on you. Play our computer game and see who's the winner.

AUGUST, 23

By visiting the tobacco field as well as communicating with the officials in the tobacco company, we got useful suggestions and inspirations. Then we designed our device and modified our "wetlab" work.

AUGUST, 25

We had a freewheeling talk with the iGEM team of Northeastern University, because of this talk, we later had opportunities to collaborate with them in experiments.

AUGHST, 26

Many suggestions that we received from other teams are of value, so we further improved our design after coming back. Besides, we built close cooperation with UESTC-China since we both used the tobacco.

OCTOBER, 7

We trained ourselves through telling others our story, and they fed back what they were interested in or what they could not figure out, and all these information helped us discover many defects in our presentation.

Human Practice Summary

We engaged with stakeholders as well as experts,and did field study etc., which as a whole helped us to design and improve our project.

We consider the safety problem and interviewed the officials about bio-safety legislation. New designs and changes occurred after we made a trip to the tobacco company. Also, we made a lot of connection with public, especially teenagers, through activities like garden party, open house day, project-related computer game and so on. What's more, we joined in some meetings with other participants.

We saw the current condition and problem, and made dialogues with professors and common people to got useful advice. Then, we made new things(like our VOC device) and confirmed one direction of our project. Through the project process, human practice helped us throw light on public issues and make efforts to solve the practical problem.


Copyright © 2017 ZJU-IGEM | Zhejiang University |