Difference between revisions of "Team:SCAU-China/Parts"

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{{SCAU-China}}
 
 
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<div class="Tan MingYang"></div>
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<div class="Li JiaDong"></div>
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<div class="Huang XinLing"></div>
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<div class="Fan ZhongZhao"></div>
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<div class="navbar-default">
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<div class="container">
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<div class="navbar-header">
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<a href="https://2018.igem.org/Team:SCAU-China" class="navbar-brand">SCAU-2018</a>
 +
</div>
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<div class="navbar">
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<ul style="float: left;" class="nav">
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<li class="dropdown">
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<a href="javascript:void(0)">TEAM</a>
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<span class="caret"></span>
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<ul class='dropdown-menu'>
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<li>
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<a href="https://2018.igem.org/Team:SCAU-China/Members">Members</a>
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</li>
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<li>
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<a href="https://2018.igem.org/Team:SCAU-China/Attributions">Attributions</a>
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</li>
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</ul>
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</li>
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<li class="dropdown">
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<a href="javascript:void(0)">PROJECT</a>
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<span class="caret"></span>
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<ul class='dropdown-menu'>
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<li><a href="https://2018.igem.org/Team:SCAU-China/ProjectOverview">Overview</a></li>
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<li><a href="https://2018.igem.org/Team:SCAU-China/Background">Background</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/Design">Design</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/SRK">Synergistic Recombination Kit</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/MM">Mathematical Model of Biological Intrinsic Regulation System</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/Type">Type II CRISPR/Cas 9 Kit</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/MOM">Method for Optimizing Microbial Cell Culture</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/Outlook">Outlook</a></li>
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<li><a href="https://2018.igem.org/Team:SCAU-China/Demonstrate">Demonstrate</a></li>
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</ul>
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<a href="javascript:void(0)">LAB WORK</a>
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<span class="caret"></span>
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<ul class='dropdown-menu'>
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<li><a href="https://2018.igem.org/Team:SCAU-China/Experiments">Experiments</a></li>
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<li><a href="https://2018.igem.org/Team:SCAU-China/Parts">Parts</a></li>
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<li><a href="https://2018.igem.org/Team:SCAU-China/Improve">Improve</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/InterLab">Interlab</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/Measurement">Measurement</a></li>
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</ul>
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</li>
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<a href="javascript:void(0)">MODEL</a>
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<span class="caret"></span>
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<li><a href="https://2018.igem.org/Team:SCAU-China/Model">Overview</a></li>
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<li><a href="https://2018.igem.org/Team:SCAU-China/Model/HAWNA">HAWNA</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/Model/PPIBoost">PPIBoost</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/Model/CultrueCondition">Cultrue Condition</a></li>
 +
</ul>
 +
</li>
 +
</ul>
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<ul style="float: right;" class="nav">
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<a href="javascript:void(0)">SAFETY</a>
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<li><a href="https://2018.igem.org/Team:SCAU-China/Safety">Safety</a></li>
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</ul>
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<li class="dropdown">
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<a href="javascript:void(0)">HUMAN PRACTICES</a>
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<span class="caret"></span>
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<ul class='dropdown-menu'>
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<li><a href="https://2018.igem.org/Team:SCAU-China/Human_Practices">Overview</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/silver">Silver</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/Integrated">Integrated</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/Public_Engagement">Public Engagement & Education</a></li>
 +
<li><a href="https://2018.igem.org/Team:SCAU-China/Collaborations">Collaborations</a></li>
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</ul>
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</li>
 +
</ul>
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</div>
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</div>
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</div>
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<div id="mainDiv">
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<div class="heart">
 +
<!-- 版心 -->
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<div id="mask">
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<!-- 半透明底板 -->
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<div class="DBoard" id="title">
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1. Parts collection
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</div>
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<div class="DBoard" id="article1">
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<!-- 展板 -->
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<p>The parts we used are listed at the table below.</p>
 +
<table class="fill">
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<thead>
 +
<tr>
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<th class="hard" style="font-size: 24px;font-weight: bold;">Part number</th>
 +
<th class="hard" style="font-size: 24px;font-weight: bold;">Type</th>
 +
<th class="hard" style="font-size: 24px;font-weight: bold;">Function description</th>
 +
<th class="hard" style="font-size: 24px;font-weight: bold;">Medal</th>
 +
</tr>
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</thead>
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<tbody>
 +
<tr>
 +
<td style="text-align: center;"><a href="http://parts.igem.org/Part:BBa_K515107:Experience">BBa_K515107 </a></td>
 +
<td style="text-align: center;">Composite</td>
 +
<td style="text-align: center;">Dendra2 cording</td>
 +
<td style="text-align: center;">Bronze</td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center;"><a href="http://parts.igem.org/Part:BBa_K2559000">BBa_K2559000</a></td>
 +
<td style="text-align: center;">Basic</td>
 +
<td style="text-align: center;">endoglucanase</td>
 +
<td style="text-align: center;">Silver </td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center;"><a href="http://parts.igem.org/Part:BBa_K2559001">BBa_K2559001</a></td>
 +
<td style="text-align: center;">Basic</td>
 +
<td style="text-align: center;">cellulose complement protein A</td>
 +
<td style="text-align: center;">Silver </td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center;"><a href="http://parts.igem.org/Part:BBa_K2559002">BBa_K2559002</a></td>
 +
<td style="text-align: center;">Basic</td>
 +
<td style="text-align: center;">β-glucosidase</td>
 +
<td style="text-align: center;">Silver</td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center;"><a href="http://parts.igem.org/Part:BBa_K2559003">BBa_K2559003</a></td>
 +
<td style="text-align: center;">Basic</td>
 +
<td style="text-align: center;">E. coli. Promoter PrplJ</td>
 +
<td style="text-align: center;">Unsubmit</td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center;"><a href="http://parts.igem.org/Part:BBa_K2559004">BBa_K2559004</a></td>
 +
<td style="text-align: center;">Basic</td>
 +
<td style="text-align: center;">E. coli. Promoter Pdapa</td>
 +
<td style="text-align: center;">Unsubmit</td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center;"><a href="http://parts.igem.org/Part:BBa_K2559011">BBa_K2559011</a></td>
 +
<td style="text-align: center;">Basic</td>
 +
<td style="text-align: center;">E. coli. Promoter PcaiF</td>
 +
<td style="text-align: center;">Unsubmit</td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center;"><a href="http://parts.igem.org/Part:BBa_K2559007">BBa_K2559007</a></td>
 +
<td style="text-align: center;">Basic</td>
 +
<td style="text-align: center;">Promoter of Heat Shock Protein Pgroel</td>
 +
<td style="text-align: center;">Unsubmit </td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center;"><a href="http://parts.igem.org/Part:BBa_K2559008">BBa_K2559008</a></td>
 +
<td style="text-align: center;">Basic</td>
 +
<td style="text-align: center;">slr0168</td>
 +
<td style="text-align: center;">Unsubmit </td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center;"><a href="http://parts.igem.org/Part:BBa_K2559012">BBa_K2559012</a></td>
 +
<td style="text-align: center;">Basic</td>
 +
<td style="text-align: center;">Promoter of RuBisCO(Prbc)</td>
 +
<td style="text-align: center;">Unsubmit </td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center;"><a href="http://parts.igem.org/Part:BBa_K2559013">BBa_K2559013</a></td>
 +
<td style="text-align: center;">Basic</td>
 +
<td style="text-align: center;">Terminator of RuBisCO(Prbc)</td>
 +
<td style="text-align: center;">Unsubmit </td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center;"><a href="http://parts.igem.org/Part:BBa_K2559005">BBa_K2559005</a></td>
 +
<td style="text-align: center;">Basic</td>
 +
<td style="text-align: center;">enhance green fluorescent protein</td>
 +
<td style="text-align: center;">Gold  </td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center;"><a href="http://parts.igem.org/Part:BBa_K2559009">BBa_K2559009</a></td>
 +
<td style="text-align: center;">Composite</td>
 +
<td style="text-align: center;">expression of eGFP</td>
 +
<td style="text-align: center;">Gold  </td>
 +
</tr>
  
 +
</tbody>
 +
</table>
 +
<p>Prepare competent cells (Escherichia coli strain DH5α) and perform transformation according to the cell measurement protocol. Meanwhile, we followed the calibration protocol supplied by the iGEM HQ to generate OD600 reference point, particle standard curve and  fluorescence standard curve to calibrate the plate reader (96 well plate) and then measured both the fluorescence and absorbance of our samples in 0-hour and 6-hour.  </p>
 +
<p>We used the setted protocol that we obtained so as to ensure everyone was doing correctly and repeatable. These can be found at <a href="https://2018.igem.org/Team:SCAU-China/Protocol">this page.</a></p>
 +
<h1>Results and discussion</h1>
 +
<p>We submitted the iGEM’s completed spreadsheet to the measurement committee before the deadline, and received confirmation that the results were shown here: 2018/7/30</p>
 +
<p>OD600 calibration</p>
 +
<table style="">
 +
<tr>
 +
<th class="no-border" style="border: 0;text-align: left; background-color: #ffffff;"></th>
 +
<th colspan="2" class="no-border" style="border: 0;text-align: left; background-color: #ffffff;">LUDOX CL-X  H2O</th>
 +
</tr>
 +
<tr>
 +
<td class="no-border" style="border: 0;text-align: left;">Replicate 1</td>
 +
<td class="blue" style=" background-color: #00ccff;text-align: left; border: 1px solid #000000;">0.059</td>
 +
<td class="blue" style=" background-color: #00ccff;text-align: left; border: 1px solid #000000;">0.039</td>
 +
</tr>
 +
<tr>
 +
<td class="no-border" style="border: 0;text-align: left;">Replicate 2</td>
 +
<td class="blue" style=" background-color: #00ccff;text-align: left; border: 1px solid #000000;">0.060</td>
 +
<td class="blue" style=" background-color: #00ccff;text-align: left; border: 1px solid #000000;">0.035</td>
 +
</tr>
 +
<tr>
 +
<td class="no-border" style="border: 0;text-align: left;">Replicate 3</td>
 +
<td class="blue" style=" background-color: #00ccff;text-align: left; border: 1px solid #000000;">0.060</td>
 +
<td class="blue" style=" background-color: #00ccff;text-align: left; border: 1px solid #000000;">0.035</td>
 +
</tr>
 +
<tr>
 +
<td class="no-border" style="border: 0;text-align: left;">Replicate 4</td>
 +
<td class="blue" style=" background-color: #00ccff;text-align: left; border: 1px solid #000000;">0.061</td>
 +
<td class="blue" style=" background-color: #00ccff;text-align: left; border: 1px solid #000000;">0.040</td>
 +
</tr>
 +
<tr>
 +
<td class="no-border" style="border: 0;text-align: left;">Arith. Mean</td>
 +
<td class="yellow" style="background-color: #ffcc00;text-align: left; border: 1px solid #000000;">0.059</td>
 +
<td class="yellow" style="background-color: #ffcc00;text-align: left; border: 1px solid #000000;">0.039</td>
 +
</tr>
 +
<tr>
 +
<td class="no-border" style="border: 0;text-align: left;">Corrected Abs600</td>
 +
<td class="yellow" style="background-color: #ffcc00;text-align: left; border: 1px solid #000000;">0.023</td>
 +
<td style="border: 0; background-color: transparent;"></td>
 +
</tr>
 +
<tr>
 +
<td class="no-border" style="border: 0;text-align: left;">Reference OD600</td>
 +
<td class="yellow" style="background-color: #ffcc00;text-align: left; border: 1px solid #000000;">0.063</td>
 +
<td style="border: 0; background-color: transparent;"></td>
 +
</tr>
 +
<tr>
 +
<td class="no-border" style="border: 0;text-align: left;">OD600/Abs600</td>
 +
<td class="yellow" style="background-color: #ffcc00;text-align: left; border: 1px solid #000000;">2.769</td>
 +
<td style="border: 0; background-color: transparent;"></td>
 +
</tr>
 +
</table>
 +
<p>Fluorescein standard curves</p>
 +
<img src="https://static.igem.org/mediawiki/2018/4/40/T--SCAU-China--Fluoresceinstandardcurves.jpg">
 +
<p>Particle standard curve</p>
 +
<img src="https://static.igem.org/mediawiki/2018/8/8d/T--SCAU-China--Particlestandardcurve.jpg">
 +
<p>Our goal is to reduce lab-to-lab variations in fluorescent intensity measurements, in order to test the variation in our sample, we caculated coefficient of variation for both colony, and compared the CV value by using different normalization method (fluorescence per OD or  fluorescence per particle)</p>
 +
<p>0th</p>
 +
<img src="https://static.igem.org/mediawiki/2018/8/8a/T--SCAU-China--0th1.jpg">
 +
<img src="https://static.igem.org/mediawiki/2018/3/37/T--SCAU-China--0th2.jpg">
 +
<p>6th</p>
 +
<img src="https://static.igem.org/mediawiki/2018/9/94/T--SCAU-China--6th1.jpg">
 +
<img src="https://static.igem.org/mediawiki/2018/a/af/T--SCAU-China--6th2.jpg">
 +
<p>After 6 hr growth, we found that the variation of each type of cells (devices) is decreased, but the CV value of devices 4 and device5 are much higher than the others. The variation of fluorescence per OD is much smaller than fluorescence per particle. Furthermore, we also found that the data from colony 2’s is usually more steady compared with colony 1.
 +
To conveniently compare the normalized value with original value, we removed the data of devices 4 and device 5, and only compare to the 6-hour’s data of colony 2 .
 +
</p>
 +
<img src="https://static.igem.org/mediawiki/2018/6/65/T--SCAU-China--interlabLast.jpg">
 +
<p>According to the graph, we found that fluorescence per particle could decrease the variation but it  depending on the devices. It seemed that usage ofe OD value to normalize is still better than particle. But one thing need to be pointed out that too steady normalized value will lose sensitivity when test the influences of some factors.  You can see this <a href="https://static.igem.org/mediawiki/2017/e/ef/T--oxford--interlab-data.xlsx">excel file</a> to check our raw and manipulated data.
 +
</p>
 +
</div>
 +
<div class="DBoard" id="title">
 +
2. Basic parts and Composite Part
 +
</div>
 +
<div class="DBoard" id="article1">
 +
<p>(1) BBa_K515107</p>
 +
<img src="https://static.igem.org/mediawiki/2018/b/b8/T--SCAU-China--parts-1.jpg"  style="margin-left: 30%" />
 +
<p>We characterized the BBa_K515107 part comprising the coding sequence BBa_K515007 which coded for Dendra2, a photoconvertible reporter protein under the control of the repressible promoter TetR (BBa_R0040) and the RBS (BBa_B0034). </p>
 +
<p>
 +
(2) BBa_K2559000/ BBa_K2559001/ BBa_K2559002
 +
</p>
 +
<img src="https://static.igem.org/mediawiki/2018/2/2f/T--SCAU-China--parts-2.jpg" style="width: 100%" />
 +
<p>
 +
These three parts were obtained from the Acetobacter xylinum bacterial cellulose synthase, as they were suspected to function together to regulate the expression and secretion of bacterial cellulose.
 +
</p>
 +
<p>
 +
The bcs Z(BBa_K2559000) is coding an endoglucanase in the process of cellulose hydrolysis, bcs H (BBa_K2559001) can express cellulose complement protein A which affects the expression levels of bcsB and bcsC, interacts with bcsD.
 +
bglX (BBa_K2559003) is a β-glucosidase.
  
−
<div class="column full_size">
+
</p>
−
<h1>Parts</h1>
+
<p>
−
<p>Each team will make new parts during iGEM and will submit them to the Registry of Standard Biological Parts. The iGEM software provides an easy way to present the parts your team has created. The <code>&lt;groupparts&gt;</code> tag (see below) will generate a table with all of the parts that your team adds to your team sandbox.</p>
+
(3) BBa_K2559003/ BBa_K2559004/ BBa_K2559011
−
<p>Remember that the goal of proper part documentation is to describe and define a part, so that it can be used without needing to refer to the primary literature. Registry users in future years should be able to read your documentation and be able to use the part successfully. Also, you should provide proper references to acknowledge previous authors and to provide for users who wish to know more.</p>
+
</p>
−
</div>
+
<img src="https://static.igem.org/mediawiki/2018/e/ec/T--SCAU-China--parts-3.jpg" style="width: 100%" />
−
 
+
<p>
−
<div class="column full_size">
+
The three parts BBa_K2559003, BBa_K2559004 and BBa_K2559011 were obtained from E. coli, and but all the promoter pRplj , pDapa and pCaif are totally different in expressional intensity(Figure1).  
−
<div class="highlight decoration_background">
+
</p>
−
<h3>Note</h3>
+
<p></p>
−
<p>Note that parts must be documented on the <a href="http://parts.igem.org/Main_Page"> Registry</a>. This page serves to <i>showcase</i> the parts you have made. Future teams and other users and are much more likely to find parts by looking in the Registry than by looking at your team wiki.</p>
+
<p></p>
−
</div>
+
<p>
−
</div>
+
(4) BBa_K2559007/ BBa_K2559008/ BBa_K2559012/ BBa_K2559013
−
 
+
</p>
−
<div class="clear extra_space"></div>
+
<img src="https://static.igem.org/mediawiki/2018/5/52/T--SCAU-China--parts-4.jpg" style="width: 100%" />
−
<div class="line_divider"></div>
+
<p>The four parts BBa_K2559007, BBa_K2559008 ,BBa_K2559012 BBa_K2559013 were used for the construction of our synergistic recombination and CRISPR/cas9 Kit</p>
−
<div class="clear extra_space"></div>
+
</div>
−
 
+
<p>School's name:SCAU</p>
−
 
+
<p>Member's name:SCAU</p>
−
 
+
<p>Designed by:SCAU</p>
−
 
+
</div>
−
 
+
</div>
−
<div class="column two_thirds_size">
+
</div>
−
<div class="highlight decoration_B_full">
+
<!-- 回到顶部按钮 -->
−
 
+
<em id="toTop"></em>
−
<h3>Adding parts to the registry</h3>
+
<script>
−
<p>You can add parts to the Registry at our <a href="http://parts.igem.org/Add_a_Part_to_the_Registry">Add a Part to the Registry</a> link.</p>
+
var dropdownBoxs = document.getElementsByClassName('dropdown');
−
 
+
var dropdownMenus = document.getElementsByClassName('dropdown-menu');
−
<p>We encourage teams to start completing documentation for their parts on the Registry as soon as you have it available. The sooner you put up your parts, the better you will remember all the details about your parts. Remember, you don't need to send us the DNA sample before you create an entry for a part on the Registry. (However, you <b>do</b> need to send us the DNA sample before the Jamboree. If you don't send us a DNA sample of a part, that part will not be eligible for awards and medal criteria.)</p>
+
for (let i = 0; i < dropdownBoxs.length; i++) {
−
<div class="button_link">
+
// console.log(dropdownBoxs[i]);
−
<a href="http://parts.igem.org/Add_a_Part_to_the_Registry">
+
// console.log(dropdownMenus[i]);
−
ADD PARTS
+
dropdownBoxs[i].index = i;
−
</a>
+
dropdownBoxs[i].onclick = function() {
−
</div>
+
var styles = document.defaultView.getComputedStyle(dropdownMenus[i]) || dropdownMenus[i].currentStyle;
−
 
+
// console.log(styles.display);
−
</div>
+
if (styles.display == 'none') {
−
</div>
+
for (let j = 0; j < dropdownBoxs.length; j++) {
−
 
+
dropdownMenus[j].style.display = 'none';
−
 
+
dropdownMenus[i].style.display = 'block';
−
 
+
}
−
<div class="column third_size">
+
} else {
−
<div class="highlight decoration_A_full">
+
for (let j = 0; j < dropdownBoxs.length; j++) {
−
<h3>Inspiration</h3>
+
dropdownMenus[j].style.display = 'none';
−
<p>We have a created  a <a href="http://parts.igem.org/Well_Documented_Parts">collection of well documented parts</a> that can help you get started.</p>
+
}
−
 
+
}
−
<p> You can also take a look at how other teams have documented their parts in their wiki:</p>
+
}
−
<ul>
+
}
−
<li><a href="https://2014.igem.org/Team:MIT/Parts"> 2014 MIT </a></li>
+
</script>
−
<li><a href="https://2014.igem.org/Team:Heidelberg/Parts"> 2014 Heidelberg</a></li>
+
<script type="text/javascript">
−
<li><a href="https://2014.igem.org/Team:Tokyo_Tech/Parts">2014 Tokyo Tech</a></li>
+
var a = function() {
−
</ul>
+
var box01 = document.getElementById("lists-DB");
−
</div>
+
var lis = document.getElementById("lists"); //获得轮播图图片盒子
−
</div>
+
var btns = document.getElementById("buttons"); //获得按钮盒子
−
 
+
var imgs = lis.getElementsByTagName("img"); //获得图片伪数组
−
 
+
btns.style.width = 24 * (imgs.length - 2) + "px";
−
<div class="clear extra_space"></div>
+
btns.style.marginLeft = -(12 * (imgs.length - 2)) + "px"; //动态赋值
−
 
+
for (var i = 0; i < imgs.length - 2; i++) {
−
 
+
//动态生成小圆点
−
 
+
var span = document.createElement("span");
−
 
+
btns.appendChild(span);
−
<div class="column full_size">
+
}
−
 
+
//轮播图正式部分
−
<h3>What information do I need to start putting my parts on the Registry?</h3>
+
var prev = document.getElementById("arr-l");
−
<p>The information needed to initially create a part on the Registry is:</p>
+
var next = document.getElementById("arr-r");
−
<ul>
+
var animated = false; //防止计时器多次被触发变量
−
<li>Part Name</li>
+
function animate(offset) {
−
<li>Part type</li>
+
var time = 300; //滚动一张图片总用时
−
<li>Creator</li>
+
var inteval = 10; //滚动一次的间隔时间
−
<li>Sequence</li>
+
var speed = offset / (time / inteval); //每次滚动滑动的像素
−
<li>Short Description (60 characters on what the DNA does)</li>
+
var left = parseInt(lis.style.left) + offset; //先计算出滚动后的left值
−
<li>Long Description (Longer description of what the DNA does)</li>
+
function go() {
−
<li>Design considerations</li>
+
animated = true; //为true代表正在运行
−
</ul>
+
//滑动函数
−
 
+
if (((speed > 0) + (parseInt(lis.style.left) < left) === 2) || ((speed < 0) + (parseInt(lis.style.left)) > left === 2)) {
−
<p>
+
lis.style.left = parseInt(lis.style.left) + speed + "px";
−
We encourage you to put up <em>much more</em> information as you gather it over the summer. If you have images, plots, characterization data and other information, please also put it up on the part page. </p>
+
setTimeout(go, inteval); //设置计时器
−
 
+
} else {
−
</div>
+
lis.style.left = left + "px";
−
 
+
if (parseInt(lis.style.left) > -1000) lis.style.left = -(1000 * (imgs.length - 2)) + "px";
−
 
+
if (parseInt(lis.style.left) < -(1000 * (imgs.length - 2))) lis.style.left = -1000 + "px";
−
<div class="clear extra_space"></div>
+
animated = false; //结束运行
−
<div class="line_divider"></div>
+
}
−
<div class="clear extra_space"></div>
+
}
−
 
+
go(); //调用函数
−
<div class="column full_size">
+
}
−
<h3>Part Table </h3>
+
prev.onclick = function() {
−
 
+
if (animated) return; //正在轮播停止
−
<p>Please include a table of all the parts your team has made during your project on this page. Remember part characterization and measurement data must go on your team part pages on the Registry. </p>
+
if (index == 0) index = spans.length - 1;
−
 
+
else index--;
−
</html>
+
animate(1000);
−
<groupparts>iGEM18 SCAU-China</groupparts>
+
showButton();
−
<html>
+
}
−
</div>
+
next.onclick = function() {
−
 
+
if (animated) return;
−
 
+
if (index == spans.length - 1) index = 0;
−
 
+
else index++;
−
 
+
animate(-1000);
 +
showButton();
 +
}
 +
var index = 0; //记录现在className为on的按钮
 +
var spans = btns.getElementsByTagName("span"); //得到btns下所有的span标签
 +
spans[0].className = "on";
 +
for (var i = 0; i < spans.length; i++) {
 +
spans[i].index = i; //自定义属性
 +
spans[i].onclick = function() {
 +
if (animated) return;
 +
if (this.className == "on") return;
 +
animate((index - this.index) * 1000);
 +
index = this.index;
 +
showButton();
 +
}
 +
}
 +
function showButton() { //显示按钮运动的函数
 +
for (var j = 0; j < spans.length; j++) { //排它思想
 +
//if(spans[j].className == "on") oldIndex = j;
 +
spans[j].className = "";
 +
spans[index].className = "on";
 +
}
 +
}
 +
var timer; //计时器变量 , 这里为什么不能为null
 +
var interval = 3000; //点击间隔时间
 +
function play() { //自动点击next函数
 +
timer = setTimeout(function() {
 +
next.onclick(); //自动点击next
 +
play(); //递归调用
 +
}, interval);
 +
}
 +
function stop() {
 +
//停止函数
 +
clearTimeout(timer);
 +
}
 +
box01.onmouseover = stop; //鼠标悬浮在轮播图上时停止
 +
box01.onmouseout = play; //鼠标离开轮播图继续
 +
play(); //先自己调用一次
 +
}
 +
a();
 +
</script>
 +
<script type="text/javascript">
 +
//封装函数js文件，方便调用
 +
function $(id) { return document.getElementById(id); } //封装获取id对象函数
 +
function show(obj) { obj.style.display = "block"; } //封装显示函数
 +
function hidden(obj) { obj.style.display = "none"; } //封装隐藏函数
 +
function scroll() {
 +
if (window.pageYOffset != null) {
 +
//ie9+ 和 其他浏览器
 +
return {
 +
top: window.pageYOffset,
 +
left: window.pageXOffset
 +
}
 +
} else if (document.compatMode == "CSS1Compat") {
 +
//非怪异浏览器：没有头部的
 +
return {
 +
top: document.documentElement.scrollTop,
 +
left: document.documentElement.scrollLeft
 +
}
 +
}
 +
return { //剩下的全部都是怪异浏览器
 +
top: document.body.scrollTop,
 +
left: document.body.scrollLeft
 +
}
 +
}
 +
var leader = 0,
 +
target = 0,
 +
timer = null; //计时器变量
 +
window.onscroll = function() {
 +
//滚动时执行的函数
 +
scroll().top > 0 ? show($("toTop")) : hidden($("toTop")); //隐藏和显示totop
 +
leader = scroll().top; //实时记录滚动距离
 +
}
 +
$("toTop").onclick = function() {
 +
//toTop被点击时执行的函数
 +
target = 0;
 +
timer = setInterval(function() {
 +
leader = leader + (target - leader) / 10;
 +
window.scrollTo(0, leader);
 +
if (leader == target) clearInterval(timer); //当到达顶端清除计时器
 +
//必须写在里面,因为在计时器执行时判断
 +
}, 20);
 +
}
 +
</script>
 
</html>
 
</html>

Revision as of 09:25, 17 October 2018

SCAU-2018
1. Parts collection

The parts we used are listed at the table below.

Part number Type Function description Medal
BBa_K515107 Composite Dendra2 cording Bronze
BBa_K2559000 Basic endoglucanase Silver
BBa_K2559001 Basic cellulose complement protein A Silver
BBa_K2559002 Basic β-glucosidase Silver
BBa_K2559003 Basic E. coli. Promoter PrplJ Unsubmit
BBa_K2559004 Basic E. coli. Promoter Pdapa Unsubmit
BBa_K2559011 Basic E. coli. Promoter PcaiF Unsubmit
BBa_K2559007 Basic Promoter of Heat Shock Protein Pgroel Unsubmit
BBa_K2559008 Basic slr0168 Unsubmit
BBa_K2559012 Basic Promoter of RuBisCO(Prbc) Unsubmit
BBa_K2559013 Basic Terminator of RuBisCO(Prbc) Unsubmit
BBa_K2559005 Basic enhance green fluorescent protein Gold
BBa_K2559009 Composite expression of eGFP Gold

Prepare competent cells (Escherichia coli strain DH5α) and perform transformation according to the cell measurement protocol. Meanwhile, we followed the calibration protocol supplied by the iGEM HQ to generate OD600 reference point, particle standard curve and fluorescence standard curve to calibrate the plate reader (96 well plate) and then measured both the fluorescence and absorbance of our samples in 0-hour and 6-hour.

We used the setted protocol that we obtained so as to ensure everyone was doing correctly and repeatable. These can be found at this page.

Results and discussion

We submitted the iGEM’s completed spreadsheet to the measurement committee before the deadline, and received confirmation that the results were shown here: 2018/7/30

OD600 calibration

LUDOX CL-X H2O
Replicate 1 0.059 0.039
Replicate 2 0.060 0.035
Replicate 3 0.060 0.035
Replicate 4 0.061 0.040
Arith. Mean 0.059 0.039
Corrected Abs600 0.023
Reference OD600 0.063
OD600/Abs600 2.769

Fluorescein standard curves

Particle standard curve

Our goal is to reduce lab-to-lab variations in fluorescent intensity measurements, in order to test the variation in our sample, we caculated coefficient of variation for both colony, and compared the CV value by using different normalization method (fluorescence per OD or fluorescence per particle)

0th

6th

After 6 hr growth, we found that the variation of each type of cells (devices) is decreased, but the CV value of devices 4 and device5 are much higher than the others. The variation of fluorescence per OD is much smaller than fluorescence per particle. Furthermore, we also found that the data from colony 2’s is usually more steady compared with colony 1. To conveniently compare the normalized value with original value, we removed the data of devices 4 and device 5, and only compare to the 6-hour’s data of colony 2 .

According to the graph, we found that fluorescence per particle could decrease the variation but it depending on the devices. It seemed that usage ofe OD value to normalize is still better than particle. But one thing need to be pointed out that too steady normalized value will lose sensitivity when test the influences of some factors. You can see this excel file to check our raw and manipulated data.

2. Basic parts and Composite Part

(1) BBa_K515107

We characterized the BBa_K515107 part comprising the coding sequence BBa_K515007 which coded for Dendra2, a photoconvertible reporter protein under the control of the repressible promoter TetR (BBa_R0040) and the RBS (BBa_B0034).

(2) BBa_K2559000/ BBa_K2559001/ BBa_K2559002

These three parts were obtained from the Acetobacter xylinum bacterial cellulose synthase, as they were suspected to function together to regulate the expression and secretion of bacterial cellulose.

The bcs Z(BBa_K2559000) is coding an endoglucanase in the process of cellulose hydrolysis, bcs H (BBa_K2559001) can express cellulose complement protein A which affects the expression levels of bcsB and bcsC, interacts with bcsD. bglX (BBa_K2559003) is a β-glucosidase.

(3) BBa_K2559003/ BBa_K2559004/ BBa_K2559011

The three parts BBa_K2559003, BBa_K2559004 and BBa_K2559011 were obtained from E. coli, and but all the promoter pRplj , pDapa and pCaif are totally different in expressional intensity(Figure1).

(4) BBa_K2559007/ BBa_K2559008/ BBa_K2559012/ BBa_K2559013

The four parts BBa_K2559007, BBa_K2559008 ,BBa_K2559012 BBa_K2559013 were used for the construction of our synergistic recombination and CRISPR/cas9 Kit

School's name:SCAU

Member's name:SCAU

Designed by:SCAU