Difference between revisions of "Team:Mingdao/Safety"

 
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{{:Team:Mingdao/navbar}}
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<html>
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<body>
  
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<p style="color:black; font-size:50px; text-align: center;">
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<div class="row background-white">
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<strong>Safety Regulation</strong></p>
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<div class="col-2 m-5">
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  <div class="path-btns" style="left:0;">
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<p style="color:black; font-size:30px;">
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<strong><u>Problem:</u></strong>
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</br>
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<p style="text-indent:2em; color:black; font-size:20px;">
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All personnel receives a tour and are informed of the rules upon entering the lab, the main rules include:</br>
+
  
−
1. No running, sudden moves within the lab.</br>
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      <div class="path">
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        </div>
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        <div id="regula-m-btn" class="path-dot" style="background-color: rgb(56, 94, 102);"></div>
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        <div class="pathWord path-word-sm">
 +
          <p>Safety Regulations</p>
 +
        </div>
 +
        </div>
  
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2. No eating within the premises of the lab.</br>
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      <div class="path">
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        <div class="pathWord path-word-sm">
 +
          <p>Lab Photos</p>
 +
        </div>
 +
        </div>
  
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3. All experiment waste must be thrown in a special bin then put through a machine for sterilization.</br>
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        <div class="pathWord path-word-sm">
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          <p>Our Goal</p>
 +
        </div>
 +
        </div>
  
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4. All pipettes must be sanitized after use.</br>
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        <div id="risk-s-btn" class="path-dot" style="top: 300px; background-color: rgb(255, 255, 255);"></div>
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          <p>Possible Risks</p>
 +
        </div>
 +
        </div>
  
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5. All equipment use must follow the using manual.</br>
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        <div class="pathWord path-word-sm">
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          <p>Risks Management</p>
 +
        </div>
 +
        </div>
 +
        </div>
 +
</div>
 +
     
 +
<div class="col-8 my-5 mr-5 ml-0">
 +
<h1 id ="d-regula">Safety Regulations </h1>
 +
<p>
 +
All personnels received a tour around the lab and were informed of the rules upon entering the lab, the main rules include:
 +
</p>
 +
<ol>
 +
<li>No running or sudden movements in the lab.</li>
 +
<li>No eating in the lab.</li>
 +
<li>All experiment waste must be thrown in a designated bin, which will then be put through a machine for sterilization.</li>
 +
<li>All pipettes must be sanitized after use.</li>
 +
<li>All equipment used must follow the user manual.</li>
 +
<li>Experiments can only be performed with gloves and lab coats on.</li>
 +
<li>All personnel must wash their hand upon entry into the laboratory and wear suitbale clothings.</li>
 +
</ol>
  
−
6. The mosquito cage must always be double sealed.</br>
+
<hr class="style-two" />
  
−
7. A bottle of ethanol must always be around when handling the mosquitoes.</br>
+
</br></br>
  
−
8. To enter the P2 lab, all personnel must be thoroughly sanitized, must wear full lab suits, remove shoes and watches, and pass through two doors.</br>
+
<p style="color: black; font-size:30px; text-align: center;" id ="d-photo">
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+
<strong><u>Photos of the LAB:</u></strong> </br>
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+
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<p style="tcolor:black; font-size:30px;">
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−
<strong><u>CO​2​ analysis:</u></strong> </br></p>
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<p style="text-indent:2em; color:black; font-size:20px;">
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−
In order to prove the seriousness , we measure the CO​2​ concentration in enclosed spaces and analysis the results.</br>
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<p style="color:black; font-size:20px; margin-left:40px;">
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−
If the concentration of CO​2​ is between :</br>
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<p style="color:black; font-size:20px; margin-left:40px;">
+
−
● 350~1000 ppm, the air is fresh</br>
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−
● 1000~2000 ppm, people feel the air turbid and start feeling drowsy</br>
+
−
● 2000~5000 ppm,people feel headache,lethargy, sluggishness, heart beat
+
−
hasten, mild nausea</br>
+
−
● above 5000 ppm, it may cause severe hypoxia, coma..</br>
+
 
</p>
 
</p>
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</br>
+
<img class="center" src="https://static.igem.org/mediawiki/2019/9/93/T--Mingdao--Safety-P1.jpg" alt="" style="width:60%">
−
 
+
−
<img class="center" src="https://static.igem.org/mediawiki/2019/d/dc/T--Mingdao--Description-P1.jpeg" alt="" style="width:60%">
+
  
 
<p style="color:black; font-size:20px; margin-left:40px; text-align: center;">
 
<p style="color:black; font-size:20px; margin-left:40px; text-align: center;">
−
  <strong>picture(1)</strong> </br>
+
  <strong>The BIOLAB at Mingdao High School.</strong> </br>
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<p style="text-indent:2em; color:black; font-size:20px; margin-left:80px;">
+
−
picture (1) is the result of our measure in a simple single room (only one person inside an enclosed space with air condition on .) According to the picture, the concentration had reached to 1000 ppm in a hour. While the subject felt asleep, it slowly declined(We speculate CO​2​ may leak out from the gaps, and the rate of venting is bigger than the rate of producing.) Until getting up next day, the speed of breathing was back to normal.</br>
+
 
</br>
 
</br>
 +
 
<p style="color:black; font-size:20px; margin-left:35px; text-align: center;">
 
<p style="color:black; font-size:20px; margin-left:35px; text-align: center;">
−
<img class="center" src="https://static.igem.org/mediawiki/2019/b/b2/T--Mingdao--Description-P2.jpeg" alt="" style="width:60%">
+
<img class="center" src="https://static.igem.org/mediawiki/2019/b/b5/T--Mingdao--Safety-P2.jpg" alt="" style="width:60%">
 
<p style="color:black; font-size:20px; margin-left:40px; text-align: center;">
 
<p style="color:black; font-size:20px; margin-left:40px; text-align: center;">
−
<strong>picture(2)</strong> </br>
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<strong>Eating and Drinking are not allowed in the Lab.</strong> </br>
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<p style="text-indent:2em; color:black; font-size:20px; margin-left:80px;">
+
−
picture(2) is the result in a regular car with four people in. In eleven minutes, the concentration in the car had increased from 2353 ppm to 4660 ppm.</br>
+
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<p style="text-indent:2em; color:black; font-size:20px; margin-left:80px;">
+
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The result shows if staying in a unventilated room for a long time will have bad influences to our health. (the more people there are, the more CO​2​ is produced.It may increase the rate of CO​2​ production and make the problem even worse.)
+
−
The problem of excessive CO​2​ concentration is indeed widely exists in our daily life and may have badly influenced our health.So it is important for people to solve these problems. </br>
+
 
</br>
 
</br>
  
−
</br>
+
<p style="color:black; font-size:20px; margin-left:35px; text-align: center;">
−
<p style="color:black; font-size:30px;">
+
<img class="center" src="https://static.igem.org/mediawiki/2019/7/75/T--Mingdao--Safety-P3.jpeg" alt="" style="width:60%">
−
<strong><u>Current method: </u></strong> </br>
+
<p style="color:black; font-size:20px; margin-left:40px; text-align: center;">
−
<p style="text-indent:2em; color:black; font-size:20px;">
+
<strong>The Laminar Flow Clean Bench in the BIOLAB, MDHS.</strong> </br>
−
we have gone to National Chung Hsing University, Taichung, Taiwan to search for current methods and come up with a better way.
+
−
</br>
+
−
<img class="center" src="https://static.igem.org/mediawiki/2019/6/65/T--Mingdao--Description-P3.jpeg" alt="" style="width:60%">
+
 
</br>
 
</br>
  
−
<p style="color:black; font-size:20px; margin-left:40px;">
+
<p style="color:black; font-size:20px; margin-left:35px; text-align: center;">
−
 
+
<img class="center" src="https://static.igem.org/mediawiki/2019/d/dc/T--Mingdao--Safety-P5.jpeg" alt="" style="width:60%">
−
<strong>Physics-Physical absorption: </strong> </br>
+
<p style="color:black; font-size:20px; margin-left:40px; text-align: center;">
−
<p style="color:black; font-size:20px; margin-left:80px;">
+
<strong>Warning Signs are posted within the LAB.</strong> </br>
−
(1) Using organic or inorganic solvents(For example: Propylene Carbonate,, and ocean water.) as absorbents</br>
+
−
(2) Using solid absorbents, such as carbon or Zeollte to absorb carbon dioxide.</br>
+
−
● Disadvantages:</br>
+
−
(1) The absorption is deteriorated due to the sulfide, causing the number of regeneration to lessen.</br>
+
−
(2) The absorption isn't effective </br>
+
−
 
+
  
 +
<p style="color:black; font-size:20px; margin-left:35px; text-align: center;">
 +
<img class="center" src="https://static.igem.org/mediawiki/2019/a/a1/T--Mingdao--Safety-P7.jpeg" alt="" style="width:60%">
 +
<p style="color:black; font-size:20px; margin-left:40px; text-align: center;">
 +
<strong>The Emergency Eyewash and Autoclave device in the BIOLAB, MDHS.</strong> </br>
  
 
</br>
 
</br>
−
<p style="color:black; font-size:20px; margin-left:40px;">
 
−
<strong>Chemistry-​Chemical absorption:</strong></br>
 
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<p style="color:black; font-size:20px; margin-left:80px;">
 
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● The chemical reaction between the absorbent and carbon dioxide is used to recycle CO​2​,and the reverse reaction is used to regenerate the absorbent.</br>
 
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● Disadvantage:There are many operational variables, making it more cumbersome to operate.</br>
 
  
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</br>
 
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<p style="color:black; font-size:20px; margin-left:40px;">
 
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<strong>Biology-biological absorption:</strong></br>
 
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<p style="color:black; font-size:20px; margin-left:80px;">
 
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● Taking advantage of photosynthesis in plants.</br>
 
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● Disadvantages:</br>
 
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(1)The efficiency is low</br>
 
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(2)The caring is time-consuming.</br>
 
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(3)The growing condition of the plants are unpredictable</br>
 
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<p style="color:black; font-size:20px; margin-left:40px;">
 
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<strong>Our approach:</strong></br>
 
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<p style="text-indent:2em; color:black; font-size:20px; margin-left:40px;">
 
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Since the solubility of CO​2​ is low in water, we combine biology and chemistry method, inserting the gene that can produce carbonic anhydrase(CA, which can increase the solubility of CO​2​ in water) into ​<I>bacillus subtilis</I>. T​hen use light and high photosynthesis efficiency algae to purify the water quality.</br>
 
 
</p>
 
</p>
  
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<p style="color:black; font-size:20px; margin-left:40px;">
+
  <h1 id ="d-goal">Our Goal</h1>
−
<strong>Comparison:</strong></br>
+
    <p>High concentration of carbon dioxide in an enclosed space can make people feel sleepy and even dizzy. While driving, drivers may fall asleep. Accidents might occur. To solve this problem, we plan to use the cyanobacteria with high photosynthetic efficiency to reduce the concentration of CO2. However, the solubility of CO2 in water is low, so we modified Bacillus spp. to produce human carbonic anhydrase (CA), which can increase the solubility of CO2 in water. Furthermore, to extend the application, we produced CYP2E1 from rabbits, which is used in xenobiotic metabolism, to eliminate benzene and chloroform. To sum up, we built up an efficient microalgae purification system to improve indoor air quality using natural enzymes such as carbonic anhydrase and xenobiotic metabolic enzymes.
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<p style="text-indent:2em; color:black; font-size:20px; margin-left:40px;">
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Physics and chemical methods must be done in lab, while our approach can be used in any place. The physics method needs very large space in the application place especially, comparing to our device, it’s light and portable.</br>
+
 +
    </p>
 +
  <h1 id ="d-risk">Possible Risks</h1>
 +
    <h6>Organisms Used</h6>
 +
      <p>E. coli DH5alpha, Bacillus subtilis 168, Synechococcus elongatus PCC 7942, Spirulina spp.</p>
 +
    <h6>Risks of Selected Organisms</h6>
 +
      <p>We use two kinds of algae: Synechococcus elongatus PCC 7942 (non-toxic lab strain, recognized as a safe organism and exists in nature) and Spirulina spp (edible food supplement), and they will not pose a threat even if it escapes from the lab.
 +
      </p>
 +
    <h6>Organisms Used</h6>
 +
      <ul><li>Escherichia coli: E.coli DH5 alpha </li>  
 +
              <li>Bacillus subtilis</li>
 +
      </ul>
 +
    <h6>Risks of Organisms Used</h6>
 +
      <p> Bacillus subtilis 168 (Generally Regarded As Safe) and E.coli strain are recognized within the Biosafety Level 1 by the US Public Health Service Guidelines and pose a subtle risk to humans, and are not known to consistently cause disease in immunocompetent adult humans, and present a minimal potential hazard to laboratory personnel and the environment.  
 +
      </p>
  
−
<table border="1"
 
−
cellpadding="10">
 
−
  <tr>
 
−
  <th>  </th>
 
−
    <th>Requirement </th>
 
−
    <th>Advantage</th>
 
−
    <th>Disadvantage</th>
 
−
  </tr>
 
−
  <tr>
 
−
  <th  align="center">Physical absorption</th>
 
−
  <td width="20%">(1) high pressure</br>(2) low temperature</td>
 
−
  <td width="30%">(1) can be used again</br>(2) the higher the pressure is, (low temperature) the higher the efficiency is</th>
 
−
  <td width="30%">(1) demand is difficult to meet</br>(2) only suitable for places with high CO2 pressure</td>
 
−
  </tr>
 
−
  <tr>
 
−
  <th  align="center">Chemical absorption</th>
 
−
  <td>chemicals</td>
 
−
  <td>high efficiency</th>
 
−
  <td>(1) can’t be used repeatedly</br>(2) by products is corrosive</td>
 
−
  </tr>
 
−
<tr>
 
−
  <th  align="center">Biological absorption</th>
 
−
  <td>(1) sunlight</br>(2) air </br>(3) water</td>
 
−
  <td>(1) environmental friendly</br>(2) low risk</th>
 
−
  <td>(1) low efficiency</br>(2) caring is required</td>
 
−
  </tr>
 
−
<tr>
 
−
  <th  align="center">Our approach</th>
 
−
  <td>electricity</td>
 
−
  <td>(1) portable</br>(2) easy to clean</br>(3) increasing the solubility of CO​2 ​in water</th>
 
−
  <td>need to clean every day</td>
 
−
  </tr>
 
−
</table>
 
−
</br>
 
  
−
<p style="color:black; font-size:30px;">
+
 
−
<strong><u>Experiment design: </u></strong> </br>
+
    <h6> Parts Used and the Risk </h6>
−
<p style="text-indent:2em; color:black; font-size:20px;">
+
    <p> <B><font size="4" color="brown"> Basic parts information</font></B></p>  
−
We use Bacillus to secrete enzymes out of the bacteria to achieve continuous production of CA enzyme without destroying the cell wall. As for algae, we choose cyanobacteria to match the appropriate temperature and illuminance to achieve better result.</br>
+
<em> <b> <font size="4" color="green">Part: BBa_K2932000</font> </b></em>
−
<p style="color:black; font-size:20px; margin-left:40px;">
+
<p>
−
<strong>Procedure:</strong></br>
+
Name: Carbonic anhydrase (KEGG: Synpcc7942_1388) / pSB1C3</br>
−
1. Insert the gene that can produce carbonic anhydrase(CA) into ​<I>bacillus subtilis</I> </br>
+
Natural function of part: an enzyme catalyzing the interconversion between carbon dioxide/water and bicarbonate/hydrogen ions
−
2. Use light and high photosynthesis efficiency algae to purify the water quality.
+
 
</br>
 
</br>
 +
Source: gDNA of Synechococcus elongatus PCC7942 </br>
 +
Usage: to increase CO2 dissolving rate </br>
 +
Species: Synechococcus elongatus PCC7942 </br>
 +
Risk Group: 1 </br>
 +
Risk to humans or environment: No
 +
Disease: No</br>
 +
      </p>
  
 +
<em> <b> <font size="4" color="green">Part: BBa_K2932001</font> </b></em>
 +
<p>
 +
Name: Carbonic anhydrase (KEGG: Synpcc7942_1447)/ pSB1C3</br>
 +
Natural function of part: an enzyme catalyzing the interconversion between carbon dioxide/water and bicarbonate/hydrogen ions
 +
</br>
 +
Source: gDNA of Synechococcus elongatus PCC7942 </br>
 +
Usage: to increase CO2 dissolving rate </br>
 +
Species: Synechococcus elongatus PCC7942 </br>
 +
Risk Group: 1 </br>
 +
Risk to humans or environment: No
 +
Disease: No</br>
 +
      </p>
 +
<em> <b> <font size="4" color="green">Part: BBa_K2932002</font> </b></em>
 +
<p>
 +
Name: Carbonic anhydrase (human CAII) / pSB1C3</br>
 +
Natural function of part: an enzyme catalyzing the interconversion between carbon dioxide/water and bicarbonate/hydrogen ions
 
</br>
 
</br>
−
<p style="color:black; font-size:30px;">
+
Source: a plasmid got from Addgene </br>
−
<strong><u>Gene cloning: </u></strong>  
+
Usage: to increase CO2 dissolving rate </br>
 +
Species: Homo sapiens (human) </br>
 +
Risk Group: No (Animals and plants have no risk group) </br>
 +
Risk to humans or environment: No
 +
Disease: No</br>
 +
      </p>
 +
<em> <b> <font size="4" color="green">Part: BBa_K2932003</font> </b></em>
 +
<p>
 +
Name: CYP2E1 / pSB1C3</br>
 +
Natural function of part: an enzyme catalyzing the xenobiotic metabolism including benzene, chloroform, etc.
 
</br>
 
</br>
−
<p style="color:black; font-size:20px; margin-left:40px;">
+
Source: DNA synthesis by Twist Bioscience company </br>
−
<strong>Part collection:</strong></br>
+
Usage: to eliminate Volatile organic compounds (VOCs) </br>
−
<p style="color:black; font-size:20px; margin-left:80px;">
+
Species: Oryctolagus cuniculus (rabbit) </br>
 +
Risk Group: No (Animals and plants have no risk group) </br>
 +
Disease: No</br>
 +
      </p>
  
−
<strong>pBS0EP​liaI ​ promoter / pBS0E </strong></br>
+
  <p> <B><font size="4" color="brown"> Composite parts information</font></B></p>  
−
<strong>BGSCID: ​<a href="http://www.bgsc.org/getdetail.php?bgscid=ECE742">ECE742</a></strong></strong></br>
+
      <p> To express enzymes of CA or CYP2E1 in Bacillus subtilis 168, we make the following composite parts using a Bacillus promoter PliaI, which is controlled by the LiaRS antibiotic-inducible two-component system.
−
PliaI is an inducible promoter from B. subtilis, and it was first developed in <a href="https://www.ncbi.nlm.nih.gov/pubmed/24295448">The Bacillus BioBrick Box 2.0​</a>.We have got this part from BGSC(​<a href="http://www.bgsc.org/">http://www.bgsc.org/​</a>).</br></br> 
+
      </p>
−
+
−
<strong>shine dalgarno / pSB1C3</strong></br>
+
−
Sequence:AAAGGAGGAA</br>
+
−
It’s within the primer we designed to replicate the CA (Synpcc1792_1388 & Sypcc1447) sequence.</br></br>
+
  
−
<strong>CA (​<a href="https://www.genome.jp/dbget-bin/www_bget?syf:Synpcc7942_1388">Synpcc7942_1388​ </a>& ​<a href="https://www.genome.jp/dbget-bin/www_bget?syf:Synpcc7942_1447">Synpcc7942_1447​</a>)</strong></br>
+
<em> <b> <font size="4" color="green">Part: BBa_K2932004</font> </b></em>
−
We have got the algae which has the CA sequence from National Chiao Tung University, Taiwan. With PCR(Polymerase chain reaction), the CA part is able to connect with the primer. To compare the different rate of dissolving carbon dioxide, we used Synpcc7942_1388 & Synpcc7942_1447 sequence respectively.</br></br>
+
<p>PliaI promoter (BBa_K823001) + RBS (BBa_B0034) + CA1388 (BBa_K2932000) + Terminator (BBa_B0015)</p>
 +
<em> <b> <font size="4" color="green">Part: BBa_K2932005</font> </b></em>
 +
<p>PliaI promoter (BBa_K823001) + RBS (BBa_B0034) + CA1447 (BBa_K2932001) + Terminator (BBa_B0015)</p>
 +
<em> <b> <font size="4" color="green">Part: BBa_K2932006</font> </b></em>
 +
<p>PliaI promoter (BBa_K823001) + RBS (BBa_B0034) + hCA (BBa_K2932002) + Terminator (BBa_B0015)</p>
 +
<em> <b> <font size="4" color="green">Part: BBa_K2932007</font> </b></em>
 +
<p>PliaI promoter (BBa_K823001) + RBS (BBa_B0034) + CYP2E1 (BBa_K2932003) + Terminator (BBa_B0015)</p>
  
−
<strong>Terminator / pSB1C3 </strong></br>
+
    <h6>Experiment Design</h6>
−
<strong>part: ​<a href="http://parts.igem.org/Part:BBa_B0015">BBa_B0015</a></strong></br>
+
      <p>We set up a system containing algae, enzyme and culture media. We then prepared the culture media with standard media of BG-11 or baking soda, liquid fertilizer, leaf fertilizer to adjust the ratio of composition and change the illuminance, temperature to find out the appropriate environment to optimize algal growth. We prepare raw lysates from Bacillus subtilis 168 producing CA or CYP2E1 by breaking down by 0.1 um glass beads, we didn’t further purify the specific enzymes and just used the total lysates as active biocatalysts. To test CO2 elimination by CA, we use indoor CO2 (500~1000 ppm) or CO2 tanks (2000 ppm and 100%) to provide the gases and analyze CO2 concentration with infrared CO2 sensor in a chemical hood. As to test chloroform, we asked NCTU-Formosa team to help in a specialized chemistry and engineering lab.
−
This is the most commonly used terminator in gene cloning with the consisting of Double terminator BBa_B0010 and BBa_B0012. We also get this part from iGEM Jamboree in its Spring 2019 Distribution-2019 Kit Plate 3.
+
      </p>
−
</br>
+
    <h6>Risks of the Experiment</h6>
−
<img class="center" src="https://static.igem.org/mediawiki/2019/c/ca/T--Mingdao--Description-P4.jpeg" alt="" style="width:60%">
+
      <p>Our experiment only compares the growth of algae under different conditions and the solubility of carbon dioxide, so it will not pose any danger. For chloroform analysis, we collaborated with experts at the NCTU-Formosa team in a specialized lab.
−
</br>
+
      </p>
−
<p style="color:black; font-size:30px;">
+
    <h6>Project Usage</h6>
 +
        <ul>
 +
          <li>A consumer product that can be bought by all people</li>
 +
          <li>A small and enclosed device</li>
 +
        </ul>
 +
    <h6>Risks of Usage</h6>
 +
      <p>The device we designed only contains two organisms and two kinds of enzyme currently: algae (edible Spirulina spp and lab strain Synechococcus elongatus PCC 7942) and enzymes (human carbonic anhydrase and rabbit CYP2E1). Even if the device is unfortunately broken, the former won't pose a threat to the environment. While the latter will be degraded naturally. Since the materials we use won't have any negative effect, safety, security or ethical risks won’t be considered.</p>
  
−
<strong><u>Expected results: </u></strong> </br>
+
  <h1 id ="d-manage">Risks Management</h1>
−
<p style=" color:black; font-size:20px; margin-left:40px;">
+
    <h6>Biosafety Level 1 <i class="fa fa-flask" aria-hidden="true"></i> </h6>
−
● reduce the concentration of CO​2​ in enclosed space efficiently</br>
+
    <h6>Expert Assistance</h6>
−
● portable</br>
+
    <p>Dr.Pei-Hong Chen will oversee us when we are doing the experiments. He has a PhD degree in Molecular Biology and is familiar with synthetic biology experiments. In addition, he is highly familiar with iGEM due to his years of experience as our school's iGEM coach. </p>
−
● moderately sized</br>
+
    <h6>Guidnace of Risk Management </h6>
−
● intelligence control is available (IoT)</br>
+
    <p>We underwent trainings on lab skills and safety by our instructor, Dr, Pei-Hong Chen, before working in the biotech laboratory. We always strictly followed the guidance of biological laboratory regulation at Taiwan given by Academia Sinica, the national academy of Taiwan. See the guide book <a href="https://biosafety.sinica.edu.tw/2e.html">here</a>.
−
● algae in the device can be used again
+
  
−
</br>
+
</p>
 +
    <h6>Topics Learned</h6>
 +
    <ul><li>Lab access and rules (including appropriate clothing, eating and drinking, etc.</li>
 +
  <li>Responsible individuals (such as lab or departmental specialist or institutional biosafety officer)</li>
 +
  <li>Differences between biosafety levels</li>
 +
  <li>Biosafety equipment (such as biosafety cabinets)</li>
 +
  <li>Good microbial techniques (such as lab practices)</li>
 +
  <li>Disinfection and sterilization</li>
 +
  <li>Emergency procedures</li>
 +
  <li>Transport rules</li>
 +
  <li>Physical biosecurity</li>
 +
  <li>Personnel biosecurity</li>
 +
  <li>Dual-use and experiments of concern</li>
 +
  <li>Data biosecurity</li>
 +
  <li>Chemicals, fire and electrical safety</li>
 +
    </ul>
 +
    <h6>Work Areas</h6>
 +
    <ul>
 +
    <li>Open bench</li>
 +
    <li>Biosafety cabinet</li>
 +
    </ul>
 +
    <h6>Risk Management Tools</h6>
 +
    <ul>
 +
    <li>Accident reporting (measures to record any accidents)</li>
 +
    <li>Personal Protective Equipment (including lab coats, gloves, eye protection, etc)</li>
 +
    </ul>
 +
  </div>
 +
</div>
 +
</div>
 +
</div>
  
−
</br>
+
<script type="text/javascript">
−
<p style="color:black; font-size:30px;">
+
    $("#regula-m-btn").click(function() {
−
<strong><u>Conclusion: </u></strong> </br>
+
      $('html, body').animate({
−
<p style="text-indent:2em; color:black; font-size:20px;">
+
          scrollTop: $("#d-regula").offset().top-70
−
In order to solve the high concentration of carbon dioxide in enclosed space, we invented our product to deal with the problem simply (without special equipments). The followings are the methods and the advantages of our device.
+
      }, 700);
−
</br>
+
    });
 +
    $("#photo-c-btn").click(function() {
 +
      $('html, body').animate({
  
−
</br>
+
          scrollTop: $("#d-photo").offset().top-70
−
<p style="color:black; font-size:30px;">
+
      }, 700);
−
<strong><u>Application: </u></strong> </br>
+
    });
−
<p style="text-indent:2em; color:black; font-size:20px;">
+
    $("#goal-c-btn").click(function() {
−
Since our device is portable, it can be used everywhere.The followings are the places that often contains high CO​2 ​concentration and can be easily improve by using our device.</br>
+
      $('html, body').animate({
−
<p style=" color:black; font-size:20px; margin-left:40px;">
+
          scrollTop: $("#d-goal").offset().top-70
−
● public transportation</br>
+
      }, 700);
−
● classroom</br>
+
    });
−
● gym</br>
+
    $("#risk-s-btn").click(function() {
−
● office</br>
+
      $('html, body').animate({
−
● cafeteria</br>
+
          scrollTop: $("#d-risk").offset().top-70
−
● house
+
      }, 700);
−
</br>
+
    });
 +
    $("#manage-t-btn").click(function() {
 +
      $('html, body').animate({
 +
          scrollTop: $("#d-manage").offset().top-70
 +
      }, 700);
 +
    });
  
−
</br>
+
    $(document).ready(function(){
−
<p style="color:black; font-size:30px;">
+
      $('.top').on('click', function(){
−
<strong><u>Signification:</u> </strong> </br>
+
        $('html, body').animate({scrollTop: '200px'}, 700);
−
<p style=" color:black; font-size:20px; margin-left:40px;">
+
      });
−
● Differ from physics or chemistry way, our product can work without special experimental equipments </br>
+
        $("#regula-m-btn").css('background-color', '#385e66');
−
● Portable and can be used everywhere </br>
+
        var scroll_pos = 0;
−
● The device operates in a circulation way and is easy to clean up. </br>
+
        $(document).scroll(function() {
−
● The algae we produce can be used in different aspects(ex.fertilizer) </br>
+
            scroll_pos = $(this).scrollTop();
−
● Improve indoor air quality in an environmentally friendly way
+
−
</br>
+
  
−
</br>
+
            d_regula_pos = $("#d-regula").offset().top -72
−
<p style="color:black; font-size:30px;">
+
            d_photo_pos = $("#d-photo").offset().top-72
−
<strong><u>Product design: </u></strong>
+
            d_goal_pos = $("#d-goal").offset().top-72
−
</br></br>
+
            d_risk_pos = $("#d-risk").offset().top-72
−
<img class="center" src="https://static.igem.org/mediawiki/2019/3/39/T--Mingdao--Description-P5.jpeg" alt="" style="width: 60%; margin-bottom: 20px;">
+
            d_manage_pos = $("#d-manage").offset().top-72
−
</br>
+
−
<p style="color:black; font-size:30px;">
+
−
<strong><u>Reference: </u></strong> </br>
+
−
<p style="text-indent:2em; color:black; font-size:20px;">
+
−
Chen, P.H., Liu, H.L., Chen, Y.J., Cheng, H., Lin, W.L., Yeh, C.H., Chang, C.H., 2012. Enhancing CO2 bio-mitigation by genetic engineering of cyanobacteria. Energy and Environmental Science 5, 8318–8327.
+
−
</br>
+
  
−
</p>
+
            // experiment
 +
            if(scroll_pos < d_photo_pos) {
 +
                $(".path-dot").css('background-color', '#fff')
 +
                $("#regula-m-btn").css('background-color', '#385e66');
 +
            }
  
−
  </div>
+
            // pest
 +
            else if(scroll_pos < d_goal_pos){
 +
              if(scroll_pos >= d_photo_pos){
 +
                $(".path-dot").css('background-color', '#fff')
 +
                $("#photo-c-btn").css('background-color', '#385e66');}
 +
            }
 +
 
 +
            // blood
 +
            else if(scroll_pos < d_risk_pos){
 +
              if(scroll_pos >= d_goal_pos){
 +
                $(".path-dot").css('background-color', '#fff')
 +
                $("#goal-c-btn").css('background-color', '#385e66');}
 +
            }
 +
 
 +
            // pathogen
 +
            else if(scroll_pos < d_manage_pos){
 +
              if(scroll_pos >= d_risk_pos){
 +
                $(".path-dot").css('background-color', '#fff')
 +
                $("#risk-s-btn").css('background-color', '#385e66');}
 +
            }
 +
 
 +
            // conclusion
 +
            else if( scroll_pos >= d_manage_pos) {
 +
                $(".path-dot").css('background-color', '#fff')
 +
                $("#manage-t-btn").css('background-color', '#385e66');}
 +
        });
 +
    });
 +
</script>
  
 
   </body>
 
   </body>
−
 
−
 
 
</html>
 
</html>
 +
{{:Team:Mingdao/test9}}

Latest revision as of 03:52, 22 October 2019

HOME
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JUDGING FORM ⇗
Responsive image

Safety Regulations

Lab Photos

Our Goal

Possible Risks

Risks Management

Safety Regulations

All personnels received a tour around the lab and were informed of the rules upon entering the lab, the main rules include:

  1. No running or sudden movements in the lab.
  2. No eating in the lab.
  3. All experiment waste must be thrown in a designated bin, which will then be put through a machine for sterilization.
  4. All pipettes must be sanitized after use.
  5. All equipment used must follow the user manual.
  6. Experiments can only be performed with gloves and lab coats on.
  7. All personnel must wash their hand upon entry into the laboratory and wear suitbale clothings.


Photos of the LAB:

The BIOLAB at Mingdao High School.

Eating and Drinking are not allowed in the Lab.

The Laminar Flow Clean Bench in the BIOLAB, MDHS.

Warning Signs are posted within the LAB.

The Emergency Eyewash and Autoclave device in the BIOLAB, MDHS.

Our Goal

High concentration of carbon dioxide in an enclosed space can make people feel sleepy and even dizzy. While driving, drivers may fall asleep. Accidents might occur. To solve this problem, we plan to use the cyanobacteria with high photosynthetic efficiency to reduce the concentration of CO2. However, the solubility of CO2 in water is low, so we modified Bacillus spp. to produce human carbonic anhydrase (CA), which can increase the solubility of CO2 in water. Furthermore, to extend the application, we produced CYP2E1 from rabbits, which is used in xenobiotic metabolism, to eliminate benzene and chloroform. To sum up, we built up an efficient microalgae purification system to improve indoor air quality using natural enzymes such as carbonic anhydrase and xenobiotic metabolic enzymes.

Possible Risks

Organisms Used

E. coli DH5alpha, Bacillus subtilis 168, Synechococcus elongatus PCC 7942, Spirulina spp.

Risks of Selected Organisms

We use two kinds of algae: Synechococcus elongatus PCC 7942 (non-toxic lab strain, recognized as a safe organism and exists in nature) and Spirulina spp (edible food supplement), and they will not pose a threat even if it escapes from the lab.

Organisms Used
  • Escherichia coli: E.coli DH5 alpha
  • Bacillus subtilis
Risks of Organisms Used

Bacillus subtilis 168 (Generally Regarded As Safe) and E.coli strain are recognized within the Biosafety Level 1 by the US Public Health Service Guidelines and pose a subtle risk to humans, and are not known to consistently cause disease in immunocompetent adult humans, and present a minimal potential hazard to laboratory personnel and the environment.

Parts Used and the Risk

Basic parts information

Part: BBa_K2932000

Name: Carbonic anhydrase (KEGG: Synpcc7942_1388) / pSB1C3
Natural function of part: an enzyme catalyzing the interconversion between carbon dioxide/water and bicarbonate/hydrogen ions
Source: gDNA of Synechococcus elongatus PCC7942
Usage: to increase CO2 dissolving rate
Species: Synechococcus elongatus PCC7942
Risk Group: 1
Risk to humans or environment: No Disease: No

Part: BBa_K2932001

Name: Carbonic anhydrase (KEGG: Synpcc7942_1447)/ pSB1C3
Natural function of part: an enzyme catalyzing the interconversion between carbon dioxide/water and bicarbonate/hydrogen ions
Source: gDNA of Synechococcus elongatus PCC7942
Usage: to increase CO2 dissolving rate
Species: Synechococcus elongatus PCC7942
Risk Group: 1
Risk to humans or environment: No Disease: No

Part: BBa_K2932002

Name: Carbonic anhydrase (human CAII) / pSB1C3
Natural function of part: an enzyme catalyzing the interconversion between carbon dioxide/water and bicarbonate/hydrogen ions
Source: a plasmid got from Addgene
Usage: to increase CO2 dissolving rate
Species: Homo sapiens (human)
Risk Group: No (Animals and plants have no risk group)
Risk to humans or environment: No Disease: No

Part: BBa_K2932003

Name: CYP2E1 / pSB1C3
Natural function of part: an enzyme catalyzing the xenobiotic metabolism including benzene, chloroform, etc.
Source: DNA synthesis by Twist Bioscience company
Usage: to eliminate Volatile organic compounds (VOCs)
Species: Oryctolagus cuniculus (rabbit)
Risk Group: No (Animals and plants have no risk group)
Disease: No

Composite parts information

To express enzymes of CA or CYP2E1 in Bacillus subtilis 168, we make the following composite parts using a Bacillus promoter PliaI, which is controlled by the LiaRS antibiotic-inducible two-component system.

Part: BBa_K2932004

PliaI promoter (BBa_K823001) + RBS (BBa_B0034) + CA1388 (BBa_K2932000) + Terminator (BBa_B0015)

Part: BBa_K2932005

PliaI promoter (BBa_K823001) + RBS (BBa_B0034) + CA1447 (BBa_K2932001) + Terminator (BBa_B0015)

Part: BBa_K2932006

PliaI promoter (BBa_K823001) + RBS (BBa_B0034) + hCA (BBa_K2932002) + Terminator (BBa_B0015)

Part: BBa_K2932007

PliaI promoter (BBa_K823001) + RBS (BBa_B0034) + CYP2E1 (BBa_K2932003) + Terminator (BBa_B0015)

Experiment Design

We set up a system containing algae, enzyme and culture media. We then prepared the culture media with standard media of BG-11 or baking soda, liquid fertilizer, leaf fertilizer to adjust the ratio of composition and change the illuminance, temperature to find out the appropriate environment to optimize algal growth. We prepare raw lysates from Bacillus subtilis 168 producing CA or CYP2E1 by breaking down by 0.1 um glass beads, we didn’t further purify the specific enzymes and just used the total lysates as active biocatalysts. To test CO2 elimination by CA, we use indoor CO2 (500~1000 ppm) or CO2 tanks (2000 ppm and 100%) to provide the gases and analyze CO2 concentration with infrared CO2 sensor in a chemical hood. As to test chloroform, we asked NCTU-Formosa team to help in a specialized chemistry and engineering lab.

Risks of the Experiment

Our experiment only compares the growth of algae under different conditions and the solubility of carbon dioxide, so it will not pose any danger. For chloroform analysis, we collaborated with experts at the NCTU-Formosa team in a specialized lab.

Project Usage
  • A consumer product that can be bought by all people
  • A small and enclosed device
Risks of Usage

The device we designed only contains two organisms and two kinds of enzyme currently: algae (edible Spirulina spp and lab strain Synechococcus elongatus PCC 7942) and enzymes (human carbonic anhydrase and rabbit CYP2E1). Even if the device is unfortunately broken, the former won't pose a threat to the environment. While the latter will be degraded naturally. Since the materials we use won't have any negative effect, safety, security or ethical risks won’t be considered.

Risks Management

Biosafety Level 1
Expert Assistance

Dr.Pei-Hong Chen will oversee us when we are doing the experiments. He has a PhD degree in Molecular Biology and is familiar with synthetic biology experiments. In addition, he is highly familiar with iGEM due to his years of experience as our school's iGEM coach.

Guidnace of Risk Management

We underwent trainings on lab skills and safety by our instructor, Dr, Pei-Hong Chen, before working in the biotech laboratory. We always strictly followed the guidance of biological laboratory regulation at Taiwan given by Academia Sinica, the national academy of Taiwan. See the guide book here.

Topics Learned
  • Lab access and rules (including appropriate clothing, eating and drinking, etc.
  • Responsible individuals (such as lab or departmental specialist or institutional biosafety officer)
  • Differences between biosafety levels
  • Biosafety equipment (such as biosafety cabinets)
  • Good microbial techniques (such as lab practices)
  • Disinfection and sterilization
  • Emergency procedures
  • Transport rules
  • Physical biosecurity
  • Personnel biosecurity
  • Dual-use and experiments of concern
  • Data biosecurity
  • Chemicals, fire and electrical safety
Work Areas
  • Open bench
  • Biosafety cabinet
Risk Management Tools
  • Accident reporting (measures to record any accidents)
  • Personal Protective Equipment (including lab coats, gloves, eye protection, etc)