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| − | <h1 hidden>About our Lab</h1>
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| − | <div class="page-header">
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| − | <h1 id="tables">About our Lab</h1>
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| − | <h2>Biosafety Level</h2><br><br>
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| − | <h3>1. What is the Safety Level of your lab?</h3><br>
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| − | <p class="style1">Level 1 (low risk)</p><br>
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| − | <h3>2.Which work areas do you use to handle biological materials? </h3><br>
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| − | <p class="style1">Open bench & Biosafety cabinet</p><br>
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| − | <h2>Biosafety Training</h2><br><br>
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| − | <h3>1.Have your team members received any safety training yet?</h3><br>
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| − | <p class="style1">Yes, we have already received safety training.</p><br>
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| − | <h3>2. briefly describe the topics that you learned about (or will learn about) in your safety training.</h3><br>
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| − | <li>• Lab access and rules (including appropriate clothing, eating and drinking, etc.) </li><br>
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| − | <li>• Responsible individuals (such as lab or departmental specialist or institutional biosafety officer) </li><br>
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| − | <li>• Differences between biosafety levels </li><br>
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| − | <li>• Biosafety equipment (such as biosafety cabinets) </li><br>
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| − | <li>• Good microbial technique (such as lab practices)</li><br>
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| − | <li>• Chemicals, fire and electrical safety</li><br>
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| − | <h3>3.Who provided your safety training? Did you get training from multiple people?</h3><br>
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| − | <p class="style1">Yes, we have already received safety training.</p><br>
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| − | <li>• Departmental specialist </li><br>
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| − | <li>• PIs/instructors</li><br>
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| − | <h2>Local rules and regulations</h2><br><br>
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| − | <h3>1.Who is responsible for the safety of biology labs at your institution? What are the guidelines for laboratory biosafety? Please give a link to these guidelines, or briefly describe them if you cannot give a link.</h3><br>
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| − | <a href="http://sbc.ecust.edu.cn/1573/list.htm">Division of laboratory & facility management is responsible for the safety of biology labs in our school.</a>
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| − | <br>The above link is the page in which some guidelines can be downloaded. (In Chinese)</p><br>
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| − | <h3>2.In your country / region, what are the laws and regulations that govern biosafety in research laboratories? Please give a link to these regulations, or briefly describe them if you cannot give a link.</h3><br>
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| − | <p class="style1"><a href="http://old.moe.gov.cn/publicfiles/business/htmlfiles/moe/s7062/201305/152275.html">Click here.</a></p><br>
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| − | <h1>Background</h1>
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| − | <p>All of the 2017 iGEM teams are invited and encouraged to participate in the Fourth International InterLaboratory Measurement Study in synthetic biology. Our team took part in this study which aimed to standardize the measurements of fluorescence in different labs. The main task was to quantify expression of GFP in common, comparable or absolute units. In our case, we measured fluorescence using plate reader.</p>
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| − | <h1>Overview</h1>
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| − | <p>Fluorescence is widely used as a proxy for promoter activity by expressing fluorescent proteins such as green fluorescent protein (GFP). Despite this is an indirect measurement, it provides a useful insight into expression levels and has significant advantage that it could be continuously monitored without disrupting cells.</p>
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| − | <p>First, we measured the standard curve by diluting supplied FITC. Using these standard curves alongside your own standard curve for FITC it is thus possible to transform your relative measurements of fluorescence into absolute measurements of GFP molecules.</p>
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| − | <p>After making the standard curve, we successfully transformed eight plasmids provided by Measurement Committee and do the cell measurement according to the protocol.</p>
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| − | <h1>Materials and methods</h1><br><br>
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| − | <p><b>plasmid used</b></p><br>
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| − | <p style="margin-left: 20px;">
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| − | Positive control<br>
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| − | Negative control<br>
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| − | Test Device 1: J23101.BCD2.E0040.B0015<br>
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| − | Test Device 2: J23106.BCD2.E0040.B0015<br>
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| − | Test Device 3: J23117.BCD2.E0040.B0015<br>
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| − | Test Device 4: J23101+I13054<br>
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| − | Test Device 5: J23106+I13054<br>
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| − | Test Device 6: J23117+I13054<br></p>
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| − | <p><b>Strain used</b></p>
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| − | <p style="margin-left: 20px;"><i>Escherichia coli</i> DH5a</p>
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| − | <p><b>Materials</b></p>
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| − | <p style="margin-left: 20px;">
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| − | LUDOX<br>
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| − | H20<br>
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| − | black flat bottom 96 well plate<br>
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| − | fluorescein<br>
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| − | 1xPBS<br>
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| − | LB(Luria Bertani) media<br>
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| − | Chloramphenicol (stock concentration 30 mg/mL dissolved in EtOH - working stock 30 ug/mL)<br>
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| − | 50ml Falcon tube<br></p>
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| − | <p><b>Machines</b></p>
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| − | Clariostar -430 -9903<br>
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| − | thermostatic shaker THZ.C<br>
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| − | electro-heating standing-temperature cultivator DHP-9082<br></p>
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| − | <p><b>Methods</b></p>
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| − | <p style="margin-left: 20px;">
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| − | <a href="">InterLab 2017 Plate Reader Protocol</a>
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| − | <h1>Results</h1>
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| − | <h3>1. Stand curve</h3><br><br>
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| − | <div class="row" align="center"> <img src="https://static.igem.org/mediawiki/2017/7/7b/Int1.png" alt="" style="width: 600px;"> </div>
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| − | <center>
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| − | <p>Table 1. Data of FITC standard curve</p></div>
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| − | <img src="https://static.igem.org/mediawiki/2017/c/cc/Int1a.png" alt="">
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| − | <center>
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| − | <p>Figure 1a. FITC standard curve（absolute）</p></div><br><br><br><br>
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| − | </center>
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| − | <div class="col-md-6" align="center">
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| − | <img src="https://static.igem.org/mediawiki/2017/a/a6/Int1b.png" alt="">
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| − | <center>
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| − | <p> Figure 1b. FITC standard curve（log）</p></div><br><br><br><br>
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| − | </center>
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| − | <h3>2. Transformation</h3>
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| − | <div class="col-md-6" align="center">
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| − | <img src="https://static.igem.org/mediawiki/2017/e/ec/Int2.png" alt="">
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| − | <center>
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| − | <p>Figure 2. Verification by Colony PCR</p></div><br><br><br><br>
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| − | <p>
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| − | P1,P2：two clones of positive control<br>
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| − | N1,N2：two clones of Negative control<br>
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| − | 1-1,1-2：two clones of Device1<br>
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| − | 2-1,2-2：two clones of Device2<br>
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| − | 3-1,3-2：two clones of Device3<br>
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| − | 4-1,4-2：two clones of Device4<br>
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| − | 5-1,5-2：two clones of Device5<br>
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| − | 6-1,6-2：two clones of Device6<br></p>
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| − | <h3>3. Cell Measurement</h3><br><br>
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| − | <div class="row" align="center"> <img src="https://static.igem.org/mediawiki/2017/8/8a/Intt2.png" alt="" style="width: 600px;"> </div>
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| − | <center>
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| − | <p>Table 2. Data of OD600（clone 1）.</p></div><br><br><br>
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| − | </center>
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| − | <div class="row" align="center"> <img src="https://static.igem.org/mediawiki/2017/d/dd/Intt3.png" alt="" style="width: 600px;"> </div>
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| − | <p>Table 3. Data of OD600（clone 2）</p></div><br><br><br>
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| − | <img src="https://static.igem.org/mediawiki/2017/2/2d/Int3.png" alt="">
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| − | <p>Figure 3.curve of OD600（clone 1）</p></div><br><br>
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| − | </center>
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| − | <div class="col-md-6" align="center">
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| − | <img src="https://static.igem.org/mediawiki/2017/8/8f/Int4.png" alt="">
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| − | <p>Figure 4.curve of OD600（clone 2）</p></div><br><br>
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| − | <div class="row" align="center"> <img src="https://static.igem.org/mediawiki/2017/2/2d/Intt4.png" alt="" style="width: 600px;"> </div>
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| − | <center>
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| − | <p>Table 4. Data of Fluorescence（clone 1）</p></div><br><br>
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| − | </center>
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| − | <div class="row" align="center"> <img src="https://static.igem.org/mediawiki/2017/c/cc/Intt5.png" alt="" style="width: 600px;"> </div>
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| − | <center>
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| − | <p>Table 5. Data of Fluorescence（clone 2）</p></div><br><br>
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| − | </center>
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| − | <img src="https://static.igem.org/mediawiki/2017/5/5e/Int5.png" alt="">
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| − | <center>
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| − | <p>Figure 5.curve of Fluorescence（clone 1）</p></div><br><br>
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| − | </center>
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| − | </div>
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| − | <div class="col-md-6" align="center">
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| − | <img src="https://static.igem.org/mediawiki/2017/6/6d/Int6.png" alt="">
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| − | <center>
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| − | <p>Figure 6.curve of Fluorescence（clone 2）</p></div><br><br>
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| − | <h1>Discussion</h1>
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| − | <specialh5>Compare (Device 1, Device 2, Device 3) or (Device 4, Device 5, Device 6)</specialh5>
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| − | <p>
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| − | 1. It is noticeable that the promoter of Device 1or4 is strongest followed by Device 2or5, Device3or6. <br>
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| − | 2. E.coli transformed by Device 1 grew slowly due to the high expression of GFP while those transformed by other Devices has similar growth rate. <br><br>
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| − | </p>
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| − | <specialh5>Compare (Device 1 and Device 4), (Device 2 and Device 5), (Device 3 and Device 6).</specialh5>
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| − | <p > RBS of Device 1,2,3 is stronger than those of Device 4,5,6.</p>
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