Difference between revisions of "Team:AHUT China/Attributions"

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<h3>★  ALERT! </h3>
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<p>This page is used by the judges to evaluate your team for the <a href="https://2018.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2018.igem.org/Judging/Awards"> award listed below</a>. </p>
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<h1>Attributions</h1>
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<p>This page is your opportunity to explain what parts of your project you did and what was done by technicians, advisers, etc. This requirement is not about literature references - these can and should be displayed throughout your wiki.
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<h3> Bronze Medal Criterion #3</h3>
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            <div class="container">
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<p> All of the work done in your project must be attributed correctly on this page. You must clearly state the work that was done by the students on your team and note any work that was done by people outside of your team, including the host labs, advisors, instructors, and individuals not on the team roster.
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Please see the <a href="https://2018.igem.org/Judging/Medals">Medals requirements page</a> for more details.</p>
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<h3> What should this page contain?</h3>
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                            <li class="nav-item active"><a class="nav-link" href="index.html">Home</a></li>
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                                <a href="#" class="nav-link dropdown-toggle" data-toggle="dropdown" role="button" aria-haspopup="true" aria-expanded="false">TEAM</a>
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<li>Clearly state what the team accomplished</li>
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                                <ul class="dropdown-menu">
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<li>General Support</li>
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                                    <li class="nav-item"><a class="nav-link" href="team.html">Team Members</a></li>
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<li>Project support and advice</li>
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                                    <li class="nav-item"><a class="nav-link" href="blog-single.html">Attribution</a>
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<li>Fundraising help and advice</li>
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<li class="nav-item"><a class="nav-link" href="blog-single.html">Sponsoring</a></li>
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<li>Lab support</li>
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                                </ul>
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<li>Difficult technique support</li>
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<li>Project advisor support</li>
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<li>Wiki support</li>
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                                <a href="#" class="nav-link dropdown-toggle" data-toggle="dropdown" role="button" aria-haspopup="true" aria-expanded="false">PROJECT</a>
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<li>Presentation coaching</li>
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<li>Human Practices support</li>
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                                    <li class="nav-item"><a class="nav-link" href="blog.html">Overview</a></li>
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<li> Thanks and acknowledgements for all other people involved in helping make a successful iGEM team</li>
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                                    <li class="nav-item"><a class="nav-link" href="blog-single.html">Background</a></li>
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<li class="nav-item"><a class="nav-link" href="blog.html">Our lab</a></li>
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                                  </li>
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                            </li>  
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<p>Tell us if your institution teaches an iGEM or synthetic biology class and when you started your project:</p>
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                                <a href="#" class="nav-link dropdown-toggle" data-toggle="dropdown" role="button" aria-haspopup="true" aria-expanded="false">PARTS</a>
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<li>Does your institution teach an iGEM or synthetic biology course?</li>
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                                <ul class="dropdown-menu">
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<li>When did you start this course?</li>
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                                    <li class="nav-item"><a class="nav-link" href="blog.html">Improved parts</a></li>
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<li>Are the syllabus and course materials freely available online?</li>
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                                    <li class="nav-item"><a class="nav-link" href="blog-single.html">New Parts</a></li>
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<li>When did you start your brainstorming?</li>
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<li>When did you start in the lab?</li>
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<li>When did you start working on  your project?</li>
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                            </li>  
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</ul>
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<li class="nav-item"><a class="nav-link" href="elements.html">Modeling</a></li>
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                                <a href="#" class="nav-link dropdown-toggle" data-toggle="dropdown" role="button" aria-haspopup="true" aria-expanded="false">Human Practices</a>
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                                    <li class="nav-item"><a class="nav-link" href="Human Practice.html">HP for Silver</a></li>
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<div class="highlight decoration_A_full">
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                                    <li class="nav-item"><a class="nav-link" href="blog-single.html">Integrated HP for gold</a></li>
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                                </ul>
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<h3>Inspiration</h3>
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                            </li>  
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<p>Take a look at what other teams have done:</p>
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                            <li class="nav-item"><a class="nav-link" href="elements.html">Collaboration</a></li>
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<ul>
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<!--                            <li class="nav-item"><a class="nav-link" href="contact.html">Contact</a></li>-->
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<li><a href="https://2011.igem.org/Team:Imperial_College_London/Team">2011 Imperial College London</a> (scroll to the bottom)</li>
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                        </ul>
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<li><a href="https://2014.igem.org/Team:Exeter/Attributions">2014 Exeter </a></li>
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                    </div>  
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<li><a href="https://2014.igem.org/Team:Melbourne/Attributions">2014 Melbourne </a></li>
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                </nav>
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<li><a href="https://2014.igem.org/Team:Valencia_Biocampus/Attributions">2014 Valencia Biocampus</a></li>
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            </div>
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                    <h2 class="page-cover-tittle">Attributions</h2>
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<div class="column two_thirds_size">
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<h3> Why is this page needed? </h3>
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                        <li><a href="index.html">Home</a></li>
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<p>The Attribution requirement helps the judges know what you did yourselves and what you had help with. We don't mind if you get help with difficult or complex techniques, but you must report what work your team did and what work was done by others.</p>
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                        <li class="active">Attributions</li>
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<p>
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                    </ol>
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For example, you might choose to work with an animal model during your project. Working with animals requires getting a license and applying far in advance to conduct certain experiments in many countries. This is difficult to achieve during the course of a summer, but much easier if you can work with a postdoc or PI who has the right licenses.</p>
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        <!--================ Latest Blog Area  =================-->
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<h3> Can we base our project on a previous one? </h3>
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<p>Yes! You can have a project based on a previous team, or based on someone else's idea, <b>as long as you state this fact very clearly and give credit for the original project.</b> </p>
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                <div align="center"> <h2 class="title_color">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Introduction</h2></div>
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                  <p>We took part in the Fifth International InterLab Measurement Study which ains to achieve the purpose of comparative measurement. The goal of this study is to obtain large amounts of data from labs across the world,to develop absolute units for measurements GFP in a plate reader to eliminate variation between labs.</p><br>
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<div align="center"><h2 class="title_color">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Materials</h2></div>
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                  <p>Plate reader: Synergy H1 (Biotek)<br>
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Plate reader plates: Corning 3603 96-Well Microplates (black plates with clear flat bottom)<br>
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Cell culture shaker: ZWYR-200D<br><br>
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Devices:<br>
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Negative control ：BBa_R0040 <br>
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Positive control ：BBa_I20270 <br>
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Device 1： BBa_J364000  <br>
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Device 2： BBa_J364001  <br>
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Device 3： BBa_J364002  <br>
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Device 4： BBa_J364007  <br>
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Device 5： BBa_J364008  <br>
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Device 6： BBa_J364009  <br>
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Note: for Device 5, we have not transformed it into DH5⍺ competent cells successfully for many times, therefore, we thank IGEM team of Nanjing University for providing the Device 5.<br>
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Calibration material: Provided in the 2018 IGEM distribution kit <br>
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Microorganism: Escherichia coli DH5⍺ strains<br>
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</p><br>
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<div align="center"><h2 class="title_color">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Methods</h2></div>
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                  <p>Following iGEM requirements, Team AHUT_China performed measurements according to these 2018 InterLab Protocols <a href="https://static.igem.org/mediawiki/2018/0/09/2018_InterLab_Plate_Reader_Protocol.pdf">https://static.igem.org/mediawiki/2018/0/09/2018_InterLab_Plate_Reader_Protocol.pdf</a> </p><br>
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<div align="center"><h2 class="title_color">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Results</h2></div>
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                  <h4>1.OD 600 reference point</h4><p>
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Using OD 600 and H2O to generate the conversion factor for the transformation later. The average of OD600 is 0.063; the correction factor (OD600/ABS600) is 3.500
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</p><br>
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  <div align="center"><img src="https://static.igem.org/mediawiki/2018/c/cb/T--AHUT_China--_LUDOX_correct_result.jpg" width="317" height="234" alt=""/></div><br><div align="center">Fig. 1 LUDOX correct value
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  </div>
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  <h4>2.Particle standard curve</h4>
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                  <p>
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We obtained the two Particle Standard Curve (normal and log scale).
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</p><br>
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  <div align="center"><img src="https://static.igem.org/mediawiki/2018/6/65/T--AHUT_China--_Fig._2_Particle_Standard_Curve.jpg" width="701" height="440" alt=""/></div><br><div align="center">Fig. 2 Particle Standard Curve
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  </div>
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<div align="center"><img src="https://static.igem.org/mediawiki/2018/7/7e/T--AHUT_China--_Fig._3_Particle_Standard_Curve_%28log_scale%29.jpg" width="701" height="440" alt=""/></div><br><div align="center">
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  Fig. 3 Particle Standard Curve (log scale)
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</div>
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<h4>3.Fluorescein standard curve</h4><p>
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Dilution serious of fluorescein were prepared and measured in a 96 well plate. A standard curve is generated to correct the cell based readings to an equivalent fluorescein concentration.<br>
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We obtained the two Fluorescein Standard Curve (normal and log scale).
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</p><br>
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<div align="center"><img src="https://static.igem.org/mediawiki/2018/2/24/T--AHUT_China--_Fig._4_Fluorescein_Standard_Curve.jpg" width="701" height="440" alt=""/></div><br><div align="center">
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  Fig. 4 Fluorescein Standard Curve
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</div>
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<div align="center"><img src="https://static.igem.org/mediawiki/2018/a/a9/T--AHUT_China--_Fig._5_Fluorescein_Standard_Curve_%28log_scale%29.jpg" width="701" height="440" alt=""/></div><br><div align="center">
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  <div align="center" >Fig. 5 Fluorescein Standard Curve (log scale) </div>
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</div>
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  <h4>4.Cell measurements</h4>
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  </ol>
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<p>&nbsp;</p><br>
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<div align="center"><img src="https://static.igem.org/mediawiki/2018/2/21/T--AHUT_China--_Fig._6_Fluorescence_Measurements_Curve_.jpg" width="732" height="492" alt=""/></div><br><div align="center">
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  <div align="center">Fig. 6 Fluorescence Measurements Curve</div>
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</div>
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    <p>Test devices 1 and 4 show high fluorescence intensity. Test devices 2 show a modest fluorescence intensity alone with positive control group, while devices3,5,6 barely show low fluorescence intensity alone with the negative control group.
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</p><br>
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<div align="center"><img src="https://static.igem.org/mediawiki/2018/3/36/T--AHUT_China--_Fig._7_Raw_OD600_Curve_.jpg" width="724" height="484" alt=""/></div><br><div align="center">
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  <div align="center">Fig. 7 Raw OD600 Curve</div>
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</div>
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    <h4>5.We obtained the Colony Forming Units per 0.1 OD600 E. coli cultures</h4>  
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<div align="center"><img src="https://static.igem.org/mediawiki/2018/f/f1/T--AHUT_China--_Fig._8_CFU_Result.jpg" width="724" height="420" alt=""/></div><br>
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    <div align="center"><img src="https://static.igem.org/mediawiki/2018/e/e5/T--AHUT_China--_Fig._8_CFU_Result1.jpg" width="732" height="492" alt=""/></div><br>
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    <div align="center">
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  <div align="center" >Fig. 8 CFU Result</div>
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  <p >&nbsp;</p>
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<div align="center"><h2 class="title_color">Discussion</h2></div>
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                <p>For Figure 3, the log graph isn’t a straight line but not 1:1 slope. In figure 6, highest fluorescence was obtained from device 4, closely followed by test device 1. Test device 2 and positive control group show a modest fluorescence intensity and device 5,6 show low fluorescence intensity, while test devices 3 barely have any fluorescence signal as well as the negative group.  
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</p>            
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<div align="center"><h2 class="title_color">Conclusion</h2></div>
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                <p>It was certainly a technical challenge to Participate in the InterLab Study. Performing the prescribed protocols with adherence to all the InterLab guidelines yielded parts of expected results, and with the completed InterLab Google Forms, confirms our team participation in this InterLab Study.
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</p>
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Revision as of 02:11, 5 October 2018

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Attributions

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     Introduction

We took part in the Fifth International InterLab Measurement Study which ains to achieve the purpose of comparative measurement. The goal of this study is to obtain large amounts of data from labs across the world,to develop absolute units for measurements GFP in a plate reader to eliminate variation between labs.


     Materials

Plate reader: Synergy H1 (Biotek)
Plate reader plates: Corning 3603 96-Well Microplates (black plates with clear flat bottom)
Cell culture shaker: ZWYR-200D

Devices:
Negative control ：BBa_R0040
Positive control ：BBa_I20270
Device 1： BBa_J364000
Device 2： BBa_J364001
Device 3： BBa_J364002
Device 4： BBa_J364007
Device 5： BBa_J364008
Device 6： BBa_J364009
Note: for Device 5, we have not transformed it into DH5⍺ competent cells successfully for many times, therefore, we thank IGEM team of Nanjing University for providing the Device 5.
Calibration material: Provided in the 2018 IGEM distribution kit
Microorganism: Escherichia coli DH5⍺ strains


     Methods

Following iGEM requirements, Team AHUT_China performed measurements according to these 2018 InterLab Protocols https://static.igem.org/mediawiki/2018/0/09/2018_InterLab_Plate_Reader_Protocol.pdf


     Results

1.OD 600 reference point

Using OD 600 and H2O to generate the conversion factor for the transformation later. The average of OD600 is 0.063; the correction factor (OD600/ABS600) is 3.500



Fig. 1 LUDOX correct value

2.Particle standard curve

We obtained the two Particle Standard Curve (normal and log scale).



Fig. 2 Particle Standard Curve

Fig. 3 Particle Standard Curve (log scale)

3.Fluorescein standard curve

Dilution serious of fluorescein were prepared and measured in a 96 well plate. A standard curve is generated to correct the cell based readings to an equivalent fluorescein concentration.
We obtained the two Fluorescein Standard Curve (normal and log scale).



Fig. 4 Fluorescein Standard Curve

Fig. 5 Fluorescein Standard Curve (log scale)

4.Cell measurements

 



Fig. 6 Fluorescence Measurements Curve

Test devices 1 and 4 show high fluorescence intensity. Test devices 2 show a modest fluorescence intensity alone with positive control group, while devices3,5,6 barely show low fluorescence intensity alone with the negative control group.



Fig. 7 Raw OD600 Curve

5.We obtained the Colony Forming Units per 0.1 OD600 E. coli cultures



Fig. 8 CFU Result

 

Discussion

For Figure 3, the log graph isn’t a straight line but not 1:1 slope. In figure 6, highest fluorescence was obtained from device 4, closely followed by test device 1. Test device 2 and positive control group show a modest fluorescence intensity and device 5,6 show low fluorescence intensity, while test devices 3 barely have any fluorescence signal as well as the negative group.

Conclusion

It was certainly a technical challenge to Participate in the InterLab Study. Performing the prescribed protocols with adherence to all the InterLab guidelines yielded parts of expected results, and with the completed InterLab Google Forms, confirms our team participation in this InterLab Study.