Difference between revisions of "Team:Makerere University/parts"

 
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<li><a  href="https://2018.igem.org/Team:Makerere_University/humanpractices">HUMAN PRACTICES</a></li>
 
<li><a  href="https://2018.igem.org/Team:Makerere_University/humanpractices">HUMAN PRACTICES</a></li>
  
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<li><a href="https://2018.igem.org/Team:Makerere_University/attributes">ATTRIBUTES</a></li>
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<li><a href="https://2018.igem.org/Team:Makerere_University/attributes">ATTRIBUTIONS</a></li>
  
 
<li><a class="active" href="https://2018.igem.org/Team:Makerere_University/parts">PARTS</a></li>
 
<li><a class="active" href="https://2018.igem.org/Team:Makerere_University/parts">PARTS</a></li>
  
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<li><a  href="https://2018.igem.org/Team:Makerere_University/awards">AWARDS  >  </a>
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<li><a  href="#">AWARDS  >  </a>
 
<ul class="submenus">
 
<ul class="submenus">
 
<li><a  href="https://2018.igem.org/Team:Makerere_University/medalcriteria">MEDAL CRITERIA</a></li
 
<li><a  href="https://2018.igem.org/Team:Makerere_University/medalcriteria">MEDAL CRITERIA</a></li
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<li><a href="https://2018.igem.org/Team:Makerere_University/judgingform">JUDGING FORM</a></li>
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<li><a href="https://igem.org/2018_Judging_Form?id=2714">JUDGING FORM</a></li>
 
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<h1>Characterization of <a style="color:yellow;" href="http://parts.igem.org/Part:BBa_I20270">BBa_I20270</a></h1><br>
  
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<h1>Human practice and Public engagement</h1>
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<p>After we participated in the interlab and did not successfully complete the lab work because of lack of one lab measurement equipment, we decided to use parts from the registry kit that was submitted to us from the headquarters to do its characterization. We characterized <a style="color:yellow;"href="http://parts.igem.org/Part:BBa_I20270">BBa_I20270</a> and below is how we did it and the results we got. However, we started the characterization process late and by the wiki freeze we will not have documented all the results as we are still in analysis.</p>
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<h2>Our Achievements in the field<h2>
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<p>IGEM Makerere consulted Professor Venard Nantulya who heads the Department of plastic waste management at Uganda Industrial Research Institute (UIRI) and he advised us that our project was a great innovation which can be conducted on a large scale to help the country get rid of large quantities of plastic waste and he assured us that UIRI is ready to provide us with support for large scale manufacture of the bacteria.</p>
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<p>The team consulted the Managing director, Kampala City Council Authority (KCCA) about the effects of plastic waste in the capital city whose concern was about the increasing disposal of plastic wastes in the water channels and he continued by advised us to go ahead with the project since it could be a great solution for the city.</p>
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<p>
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The team also engaged with one of the mostly affected suburbs in Kampala city called Bwaise where the residents were much worried about the increasing disposal of plastic wastes into the water channels which resulted into clogging that leads into flooding during rainy seasons which brings a threat of water-associated diseases like Cholera and Malaria.<br>
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There were happy and welcomed our solution and encouraged us to go ahead wit the project.  
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</p>
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<p>
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The team also had a brief engagement with the technician in charge of Environmental Health in National Environmental Management Authority (NEMA) who expressed his concern about the current methods used in degrading and recycling plastic wastes which lead to environmental pollution.<br>
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He welcomed our project and promised that they will be of support where necessary.
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</p>
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<br>
 
<br>
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<h1>Public Engagement</h1>
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<h2>Part Protocols</h2>
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<p>IGEM Makerere conducted on seminars and public presentations in synthetic biology around the university and other institutions. The Team organised weekly seminars at the College where we expressed our idea for the project and we introduced synthetic biology as a novel science which is intended to solve challenges in the environmental health for both animals and humans.<br>
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<p>Testing the IGEM part BBa-120270, a positive control for the interlab measurement study.</p><br>
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Through these seminars we were able to achieve our goal of extending synthetic biology to the public, most of the attendees welcomed this science and saw it interesting and more practical.
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<br>
 
<br>
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The team also managed to present at the 7th Annual scientific and cultural BOMA at the College Of Veterinary Medicine Animal Resources and Biosecurity (COVAB) , Makerere University which was attended by scientists from different instutions and here we were also able to showcase our OT-2 robot from Opentrons and demonstrated how it is operated.
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<h3>Procedure</h3>
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<ol>
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<li> Suspended the plasmid DNA with 10uL of distilled water, left it to stand for 5 minutes.</li>
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<li> Transformed the competent E.coli BL21 cells with 1uL of the resuspended plasmid DNA.</li>
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<li> Cultured the transformed cells on LB agar overnight at 37<sup>0</sup>C</li>
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<li> Transferred one single colony of the transformants to freshly prepared LB broth, 10mL  of LB broth</li>
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<li> Incubated for 6 hours at 37<sup>0</sup>C at 250rpm on a rotary incubator.</li>
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<li> At an OD>0.5, picked a 1mL aliquot from the cell suspension pre-induction, lysed the cells with chloroform, a drop of chloroform to a milliliter of cell suspension then vortexed to breakdown the cells.</li>
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<li> Pipetted 100uL of supernatant into a well on a 96 well plate, then determined the absorbance of the green fluorescent protein at 500nm wavelength.</li>
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<li> Induced expression for 2hours using IPTG, then picked a 1mL cell aliquot, lysed the cells with chloroform and analyzed the supernatant to determine the absorbance at 500nm wavelength.</li>
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</ol>
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<h3>Challenges</h3>
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<p>Limited information about the plasmid and gene circuit.
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</p> 
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<br>
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<p>The table below shows the results of the part characterised using our genes.</p>
 
<br>
 
<br>
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The team was also invited to attend the conference at the Uganda Virus Research Institute on the 26th of July, 2018 where we were able to present about our project and synthetic biology and how it can solve the challenges in our daily lives.
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<table style="background:transparent; color:white;">
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</p>  
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<tr><td></td><td><b>A</b></td></tr>
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<tr><td><b>1</b></td><td>0.318</td></tr>
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<tr><td><b>2</b></td><td>0.308</td></tr>
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<tr><td><b>3</b></td><td>0.308</td></tr>
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<tr><td><b>4</b></td><td>0.329</td></tr>
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</table>
 
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</div>
 
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<h1>Sponsors</h1>
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<h1 style="color:white;">Sponsors</h1>
  
 
<img id="mak" src="https://static.igem.org/mediawiki/2018/a/a2/T--Makerere_University--Mak_logo_dd.jpg">
 
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background:url("https://static.igem.org/mediawiki/2018/a/ae/T--makerere_university--Igem_logo.jpg");
 
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Latest revision as of 18:04, 17 October 2018

Home page Wiki


Characterization of BBa_I20270


After we participated in the interlab and did not successfully complete the lab work because of lack of one lab measurement equipment, we decided to use parts from the registry kit that was submitted to us from the headquarters to do its characterization. We characterized BBa_I20270 and below is how we did it and the results we got. However, we started the characterization process late and by the wiki freeze we will not have documented all the results as we are still in analysis.


Part Protocols

Testing the IGEM part BBa-120270, a positive control for the interlab measurement study.



Procedure

  1. Suspended the plasmid DNA with 10uL of distilled water, left it to stand for 5 minutes.
  2. Transformed the competent E.coli BL21 cells with 1uL of the resuspended plasmid DNA.
  3. Cultured the transformed cells on LB agar overnight at 370C
  4. Transferred one single colony of the transformants to freshly prepared LB broth, 10mL of LB broth
  5. Incubated for 6 hours at 370C at 250rpm on a rotary incubator.
  6. At an OD>0.5, picked a 1mL aliquot from the cell suspension pre-induction, lysed the cells with chloroform, a drop of chloroform to a milliliter of cell suspension then vortexed to breakdown the cells.
  7. Pipetted 100uL of supernatant into a well on a 96 well plate, then determined the absorbance of the green fluorescent protein at 500nm wavelength.
  8. Induced expression for 2hours using IPTG, then picked a 1mL cell aliquot, lysed the cells with chloroform and analyzed the supernatant to determine the absorbance at 500nm wavelength.

Challenges

Limited information about the plasmid and gene circuit.


The table below shows the results of the part characterised using our genes.


A
10.318
20.308
30.308
40.329