Difference between revisions of "Team:KUAS Korea/Collaborations"

 
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<div class="container-fluid page-heading" style="background-image: url(https://static.igem.org/mediawiki/2017/a/a5/Collaboration.jpeg)">
<div class="container-fluid page-heading" style="background-image: url(https://static.igem.org/mediawiki/2016/9/95/20160709_170210.jpg)">
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     <h3>Collaborations</h3>
     <h3>Collaborations</h3><br><br>
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<h2>Protocols for molecular biology</h2>
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<h2>Collaborations</h2>
 
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             <div class="section" id="">
 
                 <div class="slim">
 
                 <div class="slim">
  
<br><br>
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<div class="image image-full">
<h4>1. <em>E.coli</em> TSS competent cells</h4><br>
+
<img src="https://static.igem.org/mediawiki/2017/6/6d/Collaboration_intro.jpeg">
 +
</div>
 +
<p><font size=4>One of the values we can learn from iGEM is the ability to collaborate with many enthusiastic students from all over the world. This year we made cooperation with students from three different continents; Peru, the United Kingdom, and Taiwan.<font></p><br>
  
<ol>Day1 :
+
<br>
<li>Prepare 20ml of TSS buffer and 200ml of LB (Both must be autoclaved)</li>
+
<li>Autoclave centrifuge pellets</li>
+
<li>Chill the centrifuge pellet and TSS buffer in 4C refrigerator.. </li>
+
<li>Grow your desired strain of <em>E.coli</em> in 3ml LB overnight. </li>
+
</ol><p></p>
+
<ol>Day2 : <li>Dilute 2ml of the culture in 200ml LB and grow till the OD600 reaches 0.4-0.5.</li>
+
            <li>Incubate the cells on ice for few hours.</li>
+
            <li>Centrifuge the cells at 3500rpm, 4C for 15minutes.</li>
+
            <li>Remove supernatant and resuspend the cells in chilled 20ml TSS buffer. </li>
+
            <li>Aliquot the 120 ul of cells in pre-chilled 1.5ml tubes and freeze the tube in liquid nitrogen. </li>
+
            <li>Store at -80C</li>
+
</ol><br><br><br>
+
  
<h4>2. TSS Buffer</h4> <br>
 
  
<li>PEG 10% (wt/vol) : 2g</li>
+
<h4 id="exp"> 1. ColegioFDR_Peru </h4><br>
<li>DMSO 5% (vol/vol) : 1ml</li>
+
<li>20mM MgCl2 : 0.08g</li>
+
<li>in 20ml LB</li>
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<li>autoclave and chill</li>
+
<br><br><br>
+
  
<h4>3. Chemical transformation</h4><br>
+
<img src="https://static.igem.org/mediawiki/2017/a/a8/Peru.png" width="330px" height="300px" allowfullscreen>   <img src="https://static.igem.org/mediawiki/2017/6/67/Peru2.png" width="330px" height="300px" allowfullscreen>
<ol>
+
<li>Thaw competent cells on ice.</li>
+
<li>Add 1ul to 5ul of DNA into 50ul of competent cells and gently mix.</li>
+
<li>Incubate on ice for 10 to 20 minutes.</li>
+
<li>Heat shock at 42C for 1 minute.</li>
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<li>Incubate on ice for 5 minutes.</li>
+
<li>Add 200ul of LB and mix thorougly.</li>
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<li>Grow in 37C shaking incubator for 1hour.</li>
+
<li>Plate the cells on a agar plate with appropriate antibiotics.</li>
+
<li>Incubate plates at 37C overnight.</li>
+
</ol>
+
  
 +
<br><br>
 +
<p><font size=4>This year, team <strong><strong>KUAS_Korea</strong></strong> has been collaborating with <a href="https://2017.igem.org/Team:ColegioFDR_Peru"><strong>ColegioFDR_Peru</strong></a> high school iGEM team. We were delighted to have this chance, thanks to the iGEM since this collaboration was accomplished based on connections we have made through the Giant Jamboree last year. We shared our trials and errors <strong><strong>KUAS_Korea</strong></strong> have made in iGEM for past few years as well as general understandings in synthetic biology which could help them starting things up. </font></p>
  
<br><br><br>
+
<p><font size=4><strong>ColegioFDR_Peru</strong> is a high school team, and this is their second year in iGEM. This time they are aiming for synthesizing keratinase-containing <em>E.coli</em>, which degrades the feather waste that can cause the avian flu at chicken farms. It is such a brilliant project reflecting their country’s socio-contextual issues, so feel free to find more of them at their wiki. </font></p>
  
 +
<p><font size=4>Since two countries, Korea and Peru is far apart to have an actual meet-up, we have consistently communicated through e-mail, Facebook, and online Skype. The first online meet-up was held on August 14th, and at the moment both teams were still at the stage of setting basic concepts and principals of the project. We discussed each team’s projects and gave comments regarding the direction of the project and further points to elaborate. Listening from the outer perspective, especially from young active students from the other side of the world was a truly helpful and intriguing experience. </font></p>
  
 +
<p><font size=4>Besides contents of the project itself, collaboration with <strong>ColegioFDR_Peru</strong> was meaningful enough in the means of giving a real help to difficulties most of the iGEMers are facing. We went through some basic requirements for iGEM together such as project scheduling, wiki management, part submission and some other fulfilments for medal criteria, which can be often too overwhelming to manage all by themselves. This also actually helped us to look back and go through our own works and make sure that everything was properly managed to be reviewed and rewarded. </font></p>
  
 +
<p><font size=4>Overall, during the whole process of communication and content sharing, we gained valuable learnings on cooperation from their genuine thoughts and authenticity towards the project. </font></p>
  
 
+
<br>
<h4>4. Diaphorase assay</h4><br>
+
<h4 id="exp"> 2. Manchester </h4>
<ol>
+
<li>Prepare autoclaved D.W, 10mM NADH, 20mM DCPIP</li>
+
<li>Prepare your sample and controls. Purified enzymes are recommended as the sample since crude bacteria cell extract could already have reducing agents that can reduce DCPIP.</li>
+
 
+
<li><table>
+
  <tr>
+
    <th>Purified enzymes</th>
+
    <th>Xul (Depends on the concentration of your purified enzymes)</th>
+
  </tr>
+
  <tr>
+
    <td>20mM DCPIP</td>
+
    <td>1ul</td>
+
  </tr>
+
  <tr>
+
    <td>10mM NADH</td>
+
    <td>2ul</td>
+
  </tr>
+
  <tr>
+
    <td>D.W</td>
+
    <td>17-X</td>
+
  </tr>
+
  <tr>
+
    <td>Total</td>
+
    <td>20ul</td>
+
  </tr>
+
</table></li>
+
<li>Check whether the colour changes.</li>
+
</ol>
+
<br><br><br>
+
 
+
 
+
 
+
<h4>5. Induction (IPTG. L-Arabionose)</h4><br>
+
 
+
<ol>Day 1 : <li>Prepare 200ml of LB and autoclave.</li>
+
            <li>Grow your desired strain of <em>E.coli</em> in 3ml LB overnight.</li>
+
</ol>
+
<ol>     
+
Day 2 : <li>Dilute 2ml of the culture in 200ml LB and grow till the OD600 reaches 0.5~0.8.</li>
+
          <li>Put your desired inducing agent into the 200ml culture.
+
                (Final concentration : IPTG - 0.5mM, L-arabionose - 0.5%)</li>
+
          <li>Incubate the 200ml culture in 20C overnight (12hours)</li>
+
</ol>
+
<br><br><br>
+
 
+
 
+
 
+
<h4>6. Enzyme purification (Mini scale)</h4><br>
+
 
+
<p><font size=4>We followed the protocols of NI-NTA Spin Kit handbook.</font></p>
+
 
+
<p><font size=4>(https://www.qiagen.com/cn/resources/resourcedetail?id=3fc8c76d-6d21-4887-9bf8-f35f78fcc2f2&lang=en)</font></p>
+
 
+
 
+
 
+
<br><br>
+
 
+
 
+
<h2>Protocols for battery device</h2>
+
 
<div class="image image-full">
 
<div class="image image-full">
<img src="https://static.igem.org/mediawiki/2016/b/b8/20160911_191655.jpg">
+
<img src="https://static.igem.org/mediawiki/2017/0/0a/Collaboration_Man.jpeg">
 
</div>
 
</div>
                        <div class="image image-full">
+
<p><font size=4>With the team <a href="https://2017.igem.org/Team:Manchester/Collaborations"><strong>Manchester</strong></a>, our team has participated in the project of compiling legal and administrative regulations upon GMMs. In this project, 2~3 teams from each continent were allocated to answer 7 questions about the regulation (i.e., What institutional body enforces the laws regarding the use of GMMs?). Us <strong><strong>KUAS_Korea</strong></strong> have addressed the answers via information provided from Korea Biosafety Clearing House, the government agency regulating the biosafety and GMOs. Additionally, we were designated to summarize the answers to the second question; Who regulates the use of GMMs on a case-by-case basis? We anticipate that this project will comfort many teams from IGEM and other stakeholders to easily inquire concurrent trends of GMM regulations from many different nations.</font></p>
                            <img src="https://static.igem.org/mediawiki/2016/c/c2/Korea_U_Seoul_figure2.jpeg">
+
                        </div>
+
  
 
+
<p><font size=4>In the course of our investigation and comparing Korea’s regulation with other nations, we have discovered some legislation deficiencies. Thus, we decided to get in touch with the member of Congress who belongs to the biosafety committee. Further information is elaborated on the human practice page of team <strong><strong>KUAS_Korea</strong></strong>. </font></p>
<h4>1. Battery device design</h4><br>
+
 
+
 
+
      <p><font size=4>① Cut 15mL tube into enough length(about 8cm).<br>
+
      ② Make a hole at 50mL tube's cap about ①'s diameter. Make 2 of it.<br>
+
      ③ Pierce a very small hole for wire next to ②'s hole.<br>
+
  ④ Connect electrode and wire with silicon waterproof adhesive.<br>
+
             (We used 2cm*5cm carbon paper with coated back)<br>
+
    ⑤ Attach ①, ③, ④ with silicon adhesive. You should paste it very well. <br>
+
           Two 50mL tube's cap should be arranged opposite. 50mL tubes will be linked from  outside.<br>
+
    ⑥ Prepare catholyte and anolyte at 50mL tube each. <br>
+
      ⑦ Add 175mM Sodium Chloride and Agar(15g/L) in DW, and autoclave it. <br>
+
           Next, put a lid on ①'s 15mL tube and fill it with salt bridge solution.<br>
+
⑧When salt bridge solidified in enough time, connect ⑤(body of device) and ⑥(catholyte and anolyte).<br>
+
      ⑨ Link ⑧ into voltage measuring equipment.<br>
+
      ⑩ After using the device, wash it softly and autoclave it.</font></p>
+
  
 
<br>
 
<br>
<br>
+
<h4 id="exp"> 3. NCTU Formosa </h4>
 
+
<div class="image image-full">
 
+
<img src="https://static.igem.org/mediawiki/2017/d/d7/NCTU.png" width="650px" height="500px" allowfullscreen>
<h4>2. Prepare catholyte and anolyte</h4><br>
+
</div>
<p> <font size=4>      ① Make a pure culture of bacteria(MR-1, BW25113, BL21, etc) in LB solid medium  with antibiotics. You can use ampicillin for culturing <em>Shewanella oneidensis</em> MR-1.<br>
+
<p><font size=4>With the team <strong>NCTU_Formosa</strong>, we have exchanged our projects and discussed possible problems that each of our projects can have. Furthermore, we were invited to an Asia-Pacific conference hosted by the <strong>NCTU_Formosa</strong>. </font></p>
      ② Make a seed culture with 4mL LB culture medium from single colony, add  antibiotics, and culture 12 hours. <br>
+
      ③ Insert 1mL of seed culture, antibiotics in 100mL LB. Culture 24 Hours.<br>
+
      ④ Put the cell down using centrifuge(3,000 RPM, 20 minutes, and 4°C).<br>
+
      ⑤ If you need cell disruption, use sonicator(ultrasonic processor). Turn on 2 second,  turn off 10 second for 1 minute. Total time is 4 minutes. <br>
+
      ⑥ Add mediator(30μM methylene blue) and substrate. It is convenient if you prepare  mediator as a stock solution.<br>
+
      ⑦ Put 30mM Sodium ferricyanide(electron acceptor) in anolyte.</font><br>
+
</p>
+
<br><br>
+
 
+
<h4>3. Electricity analysis</h4><br>
+
<p> <font size=4>     ① We measured voltage every single minute by an electric measuring circuit using  potentiometer(1,000 ohm) and Keithley Digital Multimeter <br>
+
      ② Therefore, we can calculate current with Ohm's law(V=IR).<br>
+
      ③ Now we can get electric power with P=VI.<br>
+
      ④ So we can draw voltage, current, and electric power graph each.<br>
+
</font>
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+
 
+
 
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</p>
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</div></div></div></div></div>
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{{:Team:KUAS_Korea/Templates/Sponsors}}
 
{{:Team:KUAS_Korea/Templates/Sponsors}}

Latest revision as of 15:13, 1 November 2017

Collaborations




Collaborations

One of the values we can learn from iGEM is the ability to collaborate with many enthusiastic students from all over the world. This year we made cooperation with students from three different continents; Peru, the United Kingdom, and Taiwan.



1. ColegioFDR_Peru




This year, team KUAS_Korea has been collaborating with ColegioFDR_Peru high school iGEM team. We were delighted to have this chance, thanks to the iGEM since this collaboration was accomplished based on connections we have made through the Giant Jamboree last year. We shared our trials and errors KUAS_Korea have made in iGEM for past few years as well as general understandings in synthetic biology which could help them starting things up.

ColegioFDR_Peru is a high school team, and this is their second year in iGEM. This time they are aiming for synthesizing keratinase-containing E.coli, which degrades the feather waste that can cause the avian flu at chicken farms. It is such a brilliant project reflecting their country’s socio-contextual issues, so feel free to find more of them at their wiki.

Since two countries, Korea and Peru is far apart to have an actual meet-up, we have consistently communicated through e-mail, Facebook, and online Skype. The first online meet-up was held on August 14th, and at the moment both teams were still at the stage of setting basic concepts and principals of the project. We discussed each team’s projects and gave comments regarding the direction of the project and further points to elaborate. Listening from the outer perspective, especially from young active students from the other side of the world was a truly helpful and intriguing experience.

Besides contents of the project itself, collaboration with ColegioFDR_Peru was meaningful enough in the means of giving a real help to difficulties most of the iGEMers are facing. We went through some basic requirements for iGEM together such as project scheduling, wiki management, part submission and some other fulfilments for medal criteria, which can be often too overwhelming to manage all by themselves. This also actually helped us to look back and go through our own works and make sure that everything was properly managed to be reviewed and rewarded.

Overall, during the whole process of communication and content sharing, we gained valuable learnings on cooperation from their genuine thoughts and authenticity towards the project.


2. Manchester

With the team Manchester, our team has participated in the project of compiling legal and administrative regulations upon GMMs. In this project, 2~3 teams from each continent were allocated to answer 7 questions about the regulation (i.e., What institutional body enforces the laws regarding the use of GMMs?). Us KUAS_Korea have addressed the answers via information provided from Korea Biosafety Clearing House, the government agency regulating the biosafety and GMOs. Additionally, we were designated to summarize the answers to the second question; Who regulates the use of GMMs on a case-by-case basis? We anticipate that this project will comfort many teams from IGEM and other stakeholders to easily inquire concurrent trends of GMM regulations from many different nations.

In the course of our investigation and comparing Korea’s regulation with other nations, we have discovered some legislation deficiencies. Thus, we decided to get in touch with the member of Congress who belongs to the biosafety committee. Further information is elaborated on the human practice page of team KUAS_Korea.


3. NCTU Formosa

With the team NCTU_Formosa, we have exchanged our projects and discussed possible problems that each of our projects can have. Furthermore, we were invited to an Asia-Pacific conference hosted by the NCTU_Formosa.