Difference between revisions of "Team:SIAT-SCIE/Demonstrate"

 
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         <div id="comic">
 
         <div id="comic">
 
             <p style="font-family:'Avenir';font-size:23px">
 
             <p style="font-family:'Avenir';font-size:23px">
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                 <img src="https://static.igem.org/mediawiki/2017/a/ac/SIAT-SCIE_proof.png" alt="comic" />
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                 <img src="https://static.igem.org/mediawiki/2017/0/01/SIAT-SCIE_Demonstration.png" alt="comic" />
 
             </p>
 
             </p>
 
         </div>
 
         </div>
 
         <div id="content">
 
         <div id="content">
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             <h2 style="font-size:x-large;">Overview</h2>
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             <h2 style="font-family:'Avenir';font-size:23px">Radiation Tolerance Experiment</h2>
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            <br/>
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             <p style="font-family:'Avenir';font-size:23px">The expression is induced by different concentration of IPTG, and use a microplate reader to obtain the fluorescence, which can indicate the amount of protein expressed. Then we dilute the broth, take 100μl of diluted broth, spread them on the petri dish, to obtain the original Clone Forming Unit per 100μl diluted broth. Then take 100μl of diluted broth, spread them on the petri dish. Place the petri dish under UV for different time periods, then grow over night, to obtain the Clone Forming Unit per 100μl of diluted irradiated broth. From these experiments, we can count the survival rates of the bacteria with different amount of Dsup protein expressed and survival rates of bacteria after different amount of radiation irradiated. </p>
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             <p style="font-family:'Avenir';font-size:23px">Our project is mainly focusing on:</p>
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             <br />
 
             <br />
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             <ul class=" block">
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             <h3 style="font-family:'Avenir';font-size:23px">Experiment Report:</h3>
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                <li style="margin-left:100px;font-family:'Avenir';font-size:23px;margin-bottom:10px">The ability of CAHS proteins in increasing the survival rate of desiccated bacteria</li>
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                <li style="margin-left:100px;font-family:'Avenir';font-size:23px;margin-bottom:10px">The ability of Dsup proteins in increasing the survival rate of irradiated bacteria</li>
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                <li style="margin-left:100px;font-family:'Avenir';font-size:23px">The amount of Dsup proteins in affecting the survival rate of irradiated bacteria</li>
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            </ul>
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             <br />
 
             <br />
 +
            <h4 style="font-family:'Avenir';font-size:23px">Sample Name:</h4>
 
             <br />
 
             <br />
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             <h2 style="font-size:x-large;">Experiment & Results</h2>
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             <h6 style="font-family:'Avenir';font-size:23px">BL21-P15A-pTac-sfGFP-Dsup-IPTG+</h6>
 +
           
 +
            <h6 style="font-family:'Avenir';font-size:23px">BL21-P15A-pTac-sfGFP-Dsup-IPTG-</h6>
 
             <br />
 
             <br />
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             <h4 style="font-family:'Avenir';font-size:23px">Desiccation Tolerance Experiment</h4>
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             <h4 style="font-family:'Avenir';font-size:23px">Notice:</h4>
 
             <br />
 
             <br />
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             <p style="font-family:'Avenir';font-size:23px">After the vector construction and protein expression, we dilute the broth and spread it evenly to the growth culture, to count the Clone Forming Unit per 100μl of diluted broth. And use the same tube of broth, dehydrate it by centrifuge and desiccator. After leaving the dried broth for 48 hours in room temperature, we rehydrate it and spread it evenly to the growth culture again, and we can obtain the Clone Forming Unit per 100μl of diluted desiccated broth. By these two data, we can calculate the survival rate of the bacteria after desiccation.
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             <h6 style="font-family:'Avenir';font-size:23px">pTac-sfGFP-Dsup is Part BBa_2472007</h6>
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            </p><br /><h4 style="font-family:'Avenir';font-size:23px"><a href="https://2017.igem.org/Team:SIAT-SCIE/Results">Results:</a></h4><br />
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            <div class="block">
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            <h5 style="font-family:'Avenir';font-size:23px">Optical Density of Broth</h5>
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            <br/><img src="https://static.igem.org/mediawiki/2017/3/36/SIAT-SCIE_proof1.png" alt="Optical Density" /><br /><br /><br />
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            </div>
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            <div class="block">
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                <div class="left">
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                    <h5 style="font-family:'Avenir';font-size:23px">Clone formed before desiccation</h5>
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                    <img src="https://static.igem.org/mediawiki/2017/e/e4/SIAT-SCIE_proof2.png" alt="Clone formed before desiccation" />
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−
                </div>
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                <div class="right">
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                    <h5 style="font-family:'Avenir';font-size:23px"> Clone formed after desiccation</h5>
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                    <img src="https://static.igem.org/mediawiki/2017/e/e4/SIAT-SCIE_proof2.png"alt=" Clone formed after desiccation" /><br /><br /><br />
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                </div>
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                <img src="https://static.igem.org/mediawiki/2017/0/00/SIAT-SCIE_proof3.png" alt="whatwhatwhat">
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−
            </div><br />
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             <br />
 
             <br />
 +
            <h4 style="font-family:'Avenir';font-size:23px">Induction:</h4>
 +
            <br />
 +
            <p style="font-family:'Avenir';font-size:23px">0.5mM/L induce for 6h</p>
 +
            <br />
 +
            <h4 style="font-family:'Avenir';font-size:23px">OD</h4>
 +
            <br />
 +
 +
            <h6 style="font-family:'Avenir';font-size:23px">BL21-P15A-pTac-sfGFP-Dsup-IPTG+</h6>
 +
            <h6 style="font-family:'Avenir';font-size:23px">BL21-P15A-pTac-sfGFP-Dsup-IPTG-</h6>
 +
            <br />
 +
 +
            <h3 style="font-family:'Avenir';font-size:23px">Results</h3>
 +
            <br />
 +
            <table>
 +
                <tr>
 +
                    <th></th>
 +
                    <th style="text-align:center;font-family:'Avenir';font-size:23px">-IPTG</th>
 +
                    <th style="text-align:center;font-family:'Avenir';font-size:23px">+IPTG</th>
 +
                </tr>
 +
                <tr>
 +
                    <th style="font-family:'Avenir';font-size:23px">UV 0s</th>
 +
                    <td style="font-family:'Avenir';font-size:23px">3072</td>
 +
                    <td style="font-family:'Avenir';font-size:23px">4244</td>
 +
                </tr>
 +
                <tr>
 +
                    <th style="font-family:'Avenir';font-size:23px">UV 5s</th>
 +
                    <td style="font-family:'Avenir';font-size:23px">4/7</td>
 +
                    <td style="font-family:'Avenir';font-size:23px">15/46</td>
 +
                </tr>
 +
                <tr>
 +
                    <th style="font-family:'Avenir';font-size:23px">UV 10s</th>
 +
                    <td style="font-family:'Avenir';font-size:23px">1/7</td>
 +
                    <td style="font-family:'Avenir';font-size:23px">11/22</td>
 +
                </tr>
 +
            </table>
 +
                        <img src="https://static.igem.org/mediawiki/2017/3/3a/SIAT-SCIE_Demonstrate1.png" alt="Demostrate1" style="width:800px;margin-left:230px;"/>
 +
            <br />
 +
            <br />
 +
            <br />
 +
            <img src="https://static.igem.org/mediawiki/2017/a/a3/SIAT-SCIE_Demonstrate2.png" alt="Demostrate2" style="width:800px;margin-left:230px;"/>
 
             <br />
 
             <br />
−
            <h4 style="font-family:'Avenir';font-size:23px">Radiation Tolerance Experiment</h4><br />
 
−
            <p style="font-family:'Avenir';font-size:23px">The expression is induced by different concentration of IPTG, and use a microplate reader to obtain the fluorescence, which can indicate the amount of protein expressed. Then we dilute the broth, take 100μl of diluted broth, spread them on the growth culture, to obtain the original Clone Forming Unit per 100μl diluted broth. Then take 100μl of diluted broth, spread them on the growth culture. Place the growth culture under UV for different time periods, then grow over night, to obtain the Clone Forming Unit per 100μl of diluted irradiated broth. From these experiments, we can count the survival rates of the bacteria with different amount of Dsup protein expressed and survival rates of bacteria after different amount of radiation irradiated.
 
−
            </p>
 
−
            <br /><h4 style="font-family:'Avenir';font-size:23px"><a href="https://2017.igem.org/Team:SIAT-SCIE/Results" title="Results">Results: </a></h4><br />
 
−
            <div class="block">
 
−
                <div class="left">
 
−
                    <h5 style="font-family:'Avenir';font-size:23px">OD:</h5>
 
−
                    <img src="https://static.igem.org/mediawiki/2017/b/b6/SIAT-SCIE_proof4.jpg" alt="OD" />
 
−
                </div>
 
−
                <div class="right">
 
−
                    <h5 style="font-family:'Avenir';font-size:23px">Fluorescence</h5>
 
−
                    <img src="https://static.igem.org/mediawiki/2017/b/b6/SIAT-SCIE_proof4.jpg" alt="Fluorescence" /><br />
 
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                </div><br /><br />
 
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            </div>
 
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            <div class="block">
 
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                <div class="left">
 
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                    <h5 style="font-family:'Avenir';font-size:23px">Clone formed before irradiation</h5>
 
−
                    <img src="https://static.igem.org/mediawiki/2017/0/00/SIAT-SCIE_proof3.png" alt="Clone formed before irradiation" />
 
−
                </div>
 
−
                <div class="right">
 
−
                    <h5 style="font-family:'Avenir';font-size:23px">Clone formed after irradiation</h5>
 
−
                    <img src="https://static.igem.org/mediawiki/2017/0/00/SIAT-SCIE_proof3.png" alt=" Clone formed after irradiation" /><br />
 
−
                </div>
 
−
            </div>
 
−
            <br/>
 
−
            <br/>
 
−
            <h2 style="font-size:x-large;">Conclusion:</h2><br/>
 
 
              
 
              
−
             <p style="font-family:'Avenir';font-size:23px">Now, we are still doing our experiment, so it is hard to say whether our theory works. Let us pray that the paper is true and we did not make any mistake.
+
            <h4 style="font-family:'Avenir';font-size:23px">Survival Rate:</h4>
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             </p>
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            <table>
−
        </div>
+
                <tr>
 +
                    <th></th>
 +
                    <th colspan="2" style="font-family:'Avenir';font-size:23px">-IPTG</th>
 +
                    <th colspan="2" style="font-family:'Avenir';font-size:23px">+IPTG</th>
 +
                </tr>
 +
                <tr>
 +
                    <th></th>
 +
                    <th style="font-family:'Avenir';font-size:23px">Dish 1</th>
 +
                    <th style="font-family:'Avenir';font-size:23px">Dish 2</th>
 +
                    <th style="font-family:'Avenir';font-size:23px">Dish 1</th>
 +
                    <th style="font-family:'Avenir';font-size:23px">Dish 2</th>
 +
                </tr>
 +
                <tr>
 +
                    <th style="font-family:'Avenir';font-size:23px">UV 5s</th>
 +
                    <td style="font-family:'Avenir';font-size:23px">0.13%</td>
 +
                    <td style="font-family:'Avenir';font-size:23px">0.23%</td>
 +
                    <td style="font-family:'Avenir';font-size:23px">0.35%</td>
 +
                    <td style="font-family:'Avenir';font-size:23px">1.08%</td>
 +
                </tr>
 +
                <tr>
 +
                    <th style="font-family:'Avenir';font-size:23px">UV 10s</th>
 +
                    <td style="font-family:'Avenir';font-size:23px">0.03%</td>
 +
                    <td style="font-family:'Avenir';font-size:23px">0.23%</td>
 +
                    <td style="font-family:'Avenir';font-size:23px">0.26%</td>
 +
                    <td style="font-family:'Avenir';font-size:23px">0.52%</td>
 +
                </tr>
 +
            </table>
 +
 
 +
               
 +
         
 +
            <br />
 +
 
 +
 
 +
            <h2 style="font-family:'Avenir';font-size:23px">Conclusion:</h2>
 +
             <p style="font-family:'Avenir';font-size:23px">We can see the survival rate of the ones with IPTG added, is significantly higher than the ones without IPTG. We can say that the presence of Dsup increase the survival rate of bacteria, after the bacteria is irradiated. So, Dsup can protect organism under radiation conditions.</p>
 +
             <br />
 +
 
 +
            <p style="font-family:'Avenir';font-size:23px">Further analysis of the data is explained on <a href="https://2017.igem.org/Team:SIAT-SCIE/Model" alt="modeling">modeling.</a> <p>
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Latest revision as of 04:00, 2 November 2017

comic

Radiation Tolerance Experiment

The expression is induced by different concentration of IPTG, and use a microplate reader to obtain the fluorescence, which can indicate the amount of protein expressed. Then we dilute the broth, take 100μl of diluted broth, spread them on the petri dish, to obtain the original Clone Forming Unit per 100μl diluted broth. Then take 100μl of diluted broth, spread them on the petri dish. Place the petri dish under UV for different time periods, then grow over night, to obtain the Clone Forming Unit per 100μl of diluted irradiated broth. From these experiments, we can count the survival rates of the bacteria with different amount of Dsup protein expressed and survival rates of bacteria after different amount of radiation irradiated.


Experiment Report:


Sample Name:


BL21-P15A-pTac-sfGFP-Dsup-IPTG+
BL21-P15A-pTac-sfGFP-Dsup-IPTG-

Notice:


pTac-sfGFP-Dsup is Part BBa_2472007

Induction:


0.5mM/L induce for 6h


OD


BL21-P15A-pTac-sfGFP-Dsup-IPTG+
BL21-P15A-pTac-sfGFP-Dsup-IPTG-

Results


-IPTG +IPTG
UV 0s 3072 4244
UV 5s 4/7 15/46
UV 10s 1/7 11/22
Demostrate1


Demostrate2

Survival Rate:

-IPTG +IPTG
Dish 1 Dish 2 Dish 1 Dish 2
UV 5s 0.13% 0.23% 0.35% 1.08%
UV 10s 0.03% 0.23% 0.26% 0.52%

Conclusion:

We can see the survival rate of the ones with IPTG added, is significantly higher than the ones without IPTG. We can say that the presence of Dsup increase the survival rate of bacteria, after the bacteria is irradiated. So, Dsup can protect organism under radiation conditions.


Further analysis of the data is explained on modeling.

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