Difference between revisions of "Team:Tacoma RAINmakers/Human Practices"

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{{Tacoma_RAINmakers}}
 
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<p>This page is used by the judges to evaluate your team for the <a href="https://2018.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2018.igem.org/Judging/Awards"> award listed below</a>. </p>
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<p> Delete this box in order to be evaluated for this medal criterion and/or award. See more information at <a href="https://2018.igem.org/Judging/Pages_for_Awards"> Instructions for Pages for awards</a>.</p>
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              <li><a href="https://2016.igem.org/Team:Imperial_College/Proof">Proof</a></li>
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              <li><a href="https://2016.igem.org/Team:Imperial_College/Basic_Part">Basic Parts</a></li>
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              <li><a href="https://2016.igem.org/Team:Imperial_College/Composite_Part">Composite Parts</a></li>
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              <li><a href="https://2016.igem.org/Team:Imperial_College/Model">Overview</a></li>
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              <li><a href="https://2016.igem.org/Team:Imperial_College/SingleCell">Single Cell Model</a></li>
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              <li><a href="https://2016.igem.org/Team:Imperial_College/GRO">Population Model</a></li>
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            <a class="dropdown-toggle active" href="https://2016.igem.org/Team:Imperial_College/Software">Software Tools </a></li>
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              <li><a href="https://2016.igem.org/Team:Imperial_College/Notebook">Lab book</a></li>
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              <li><a href="https://2016.igem.org/Team:Imperial_College/Medals">Medal Criteria</a></li>
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              <li><a href="https://2016.igem.org/Team:Imperial_College/BrainStorm">Brainstorm</a></li>
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            <a class="dropdown-toggle active" href="https://2016.igem.org/Team:Imperial_College/Human_Practices">Human Practices <span class="caret"></span></a>
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            <ul class="dropdown-menu">
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              <li><a href="https://2016.igem.org/Team:Imperial_College/Human_Practices">Human Practices</a></li>
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              <li><a href="https://2016.igem.org/Team:Imperial_College/Integrated_Practices">Integrated Practices</a></li>
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              <li><a href="https://2016.igem.org/Team:Imperial_College/Engagement">Public Engagement</a></li>
 +
              <li><a href="https://2016.igem.org/Team:Imperial_College/Collaborations">Collaborations</a></li>
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              <li><a href="https://2016.igem.org/Team:Imperial_College/Team">Team</a></li>
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              <li><a href="https://2016.igem.org/Team:Imperial_College/Attributions">Attributions</a></li>
 +
 
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          </li>
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        </ul>
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    <!--
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-->
  
  
−
<div class="clear"></div>
 
  
 +
<div class="col-lg-10 col-centered">
 +
    <!--
 +
<p>On this page, the ecolibrium team would like to share with you the protocols that we have been using over the summer. This library of protocols has been developed alongside our supervisors for the purpose of this study. Here you can find the exact methods we use to generate our data and results. We share them in the interest of reproducibility.<br><br></p>
 +
-->
 +
    <h2> Integrated Human Practices <br><br></h2>
 +
   
 +
    <p>As our team’s goal was to create a product for the community, integrating human practices was vitally important.
 +
<br>
 +
Click the titles below to see more!
  
 +
<br><br></p>
 +
   
 +
<div class="panel-group" id="accordion" role="tablist" aria-multiselectable="true">
 +
            <div class="panel panel-default">
 +
                <div class="panel-heading" role="tab" id="P1">
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                    <h4 class="panel-title">
 +
                    <a role="button" data-toggle="collapse" data-parent="#accordion" href="#P1-collapse" aria-expanded="false" aria-controls="P1-collapse">
 +
<div>
 +
                    <div class="col-md-11">Sue's Tech Kitchen</div><div class="col-md-1"><i class="fa fa-arrow-down fa-10" aria-hidden="true"></i></div>
 +
</div>
 +
                    </a>
 +
                    </h4>
  
−
<div class="column full_size">
+
                </div>
 +
                <div id="P1-collapse" class="panel-collapse collapse" role="tabpanel" aria-labelledby="P1">
 +
                    <div class="panel-body">
 +
 
 +
<!-- <p>
 +
<b>Materials:</b><br>
 +
LB broth<br>
 +
Ice<br>
 +
Selection plates<br>
 +
<br>
 +
<b>Methods:</b><br>
 +
<ol style="font-size:16px;">
 +
<li>Thaw 50µL competent E. coli cells on ice for 10 minutes<br></li>
 +
<li>Add:
 +
<ul style="font-size:16px;">
 +
<li>5-10 µl DNA from a ligation reaction mix or </li>
 +
<li>10-100ng DNA of a known plasmid </li>
 +
</ul>
 +
</li>
 +
<li>Carefully flick the tube 4-5 times to mix cells and DNA. <b>Do not vortex.</b></li>
 +
<li>Place the mixture on ice for 30 minutes. <b>Do not mix.</b></li>
 +
<li>Heat shock at exactly 42°C for exactly 30 seconds. <b>Do not mix.</b></li>
 +
<li>Place on ice for 5 minutes. <b>Do not mix.</b></li>
 +
<li>Pipette 950 µl of room temperature SOC or LB media into the mixture.</li>
 +
<li>Incubate at 37°C and 200-250 rpm for 60 minutes.</li>
 +
<li>Mix the cells thoroughly by flicking the tube and inverting.</li>
 +
<li>Spread:
 +
<ul style="font-size:16px;">
 +
<li>For ligation reaction DNA: 100µl of each transformation reaction onto a selection plate. For the rest of 900 µL:
 +
<ol style="font-size:16px;">
 +
<li>Pellet cells at 8000rpm for 3 minutes</li>
 +
<li>Remove and dispense 600 µL of supernatant </li>
 +
<li>Re-suspend cells by light vortexing</li>
 +
<li>Plate resuspended cells as above</li>
 +
</ol></li>
 +
 
 +
<li>For known plasmid: 10 &amp; 100 µL of each transformation reaction onto a selection plate. For the rest of 890 µL:
 +
<ol style="font-size:16px;">
 +
<li>Pellet cells at 8000rpm for 3 minutes</li>
 +
<li>Remove and dispense 600 µL of supernatant </li>
 +
<li>Re-suspend cells by light vortexing</li>
 +
<li>Plate resuspended cells as above</li>
 +
</ol></li>
 +
</ul>
 +
<li>Incubate overnight at 37°C with plates upside down.</li>
 +
</ol>
 +
                       
 +
</p>
 +
-->
 +
                      <p>
 +
                      Sue’s Tech Kitchen, the creation of Randi Zuckerberg, is a nationwide event dedicated to getting people excited about science through food. It’s known for its’ 3D printed smores and miracle berries, but it also serves as an opportunity for local groups to share their projects and visions with the community. We held a booth at the event in Tacoma, and had a blast telling our community all about our project. We created a mockup of our project with pH strips to represent the sensor, and solutions of a known acidity to represent water samples from around Puget Sound. Kids loved the idea of putting what looked like paper, into what looked like water, and seeing the paper change colors before their eyes. Adults were more excited about the idea of getting to test their own water and get accurate results that were easy to read. On a whiteboard, we set up a magnetic puzzle of our plasmid parts. Guests got to put together the puzzle as fast as they could, and the best time of each day won a cool water bottle. We met some incredible people, including a woman who helped create the map of arsenic contamination caused by the smelter. We had many people ask to buy test strips, but since the sensor isn’t complete, they settled for signing up for the email list. The event gave us great insight on what matters to the community, and we even got ideas on how to expand the sensor in the future!             
 +
                        </p>
 +
                       
 +
                       
 +
<!--                      <div class="heidirow">
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 +
  </div>
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  <div class="heidicolumn">
 +
    <img class = "heidisize" src="https://static.igem.org/mediawiki/2018/d/dc/T--Tacoma_RAINmakers--BiocampGroup.jpg" alt="Biocamp" style="width:100%">
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 +
    <img class = "heidisize" src="https://static.igem.org/mediawiki/2018/1/1f/T--Tacoma_RAINmakers--BiocampGroupPhoto.jpg" alt="Biocamp" style="width:100%">
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<div>
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                    <div class="col-md-11">Interview with Greg Tanbara</div><div class="col-md-1"><i class="fa fa-arrow-down fa-10" aria-hidden="true"></i></div>
 +
</div>
 +
                    </a>
 +
                    </h4>
  
−
<h1>Human Practices</h1>
+
                </div>
 +
                <div id="P2-collapse" class="panel-collapse collapse" role="tabpanel" aria-labelledby="P2">
 +
                    <div class="panel-body">
 +
    <!--
 
<p>
 
<p>
−
At iGEM we believe societal considerations should be upfront and integrated throughout the design and execution of synthetic biology projects. “Human Practices” refers to iGEM teams’ efforts to actively consider how the world affects their work and the work affects the world. Through your Human Practices activities, your team should demonstrate how you have thought carefully and creatively about whether your project is responsible and good for the world. We invite you to explore issues relating (but not limited) to the ethics, safety, security, and sustainability of your project, and to show how this exploration feeds back into your project purpose, design and execution.
+
<b>Materials:</b><br>
 +
5 ml LB broth<br>
 +
5 μl antibiotic<br>
 +
Loops<br>
 +
12 ml culture tube<br>
 +
<br>
 +
<b>Methods:</b><br>
 +
Overnight cultures were prepared under sterile conditions using a Bunsen burner<br>
 +
<ol style="font-size:16px;">
 +
<li>Add 5 ml liquid LB media into 12 ml culture tubes</li>
 +
<li>Add 5 μl of appropriate antibiotic into the broth</li>
 +
<li>Using the loop, pick a single colony and inoculate the cultures by dipping the loop into the LB broth</li>
 +
<li>Seal the tubes and incubate overnight at 37°C shaking at 200-250 rpm</li>
 +
</ol>
 
</p>
 
</p>
 +
-->
 +
                        <p>
 +
                        To continue familiarizing ourselves with problems caused by arsenic contamination in the environment, we turned to our local health department. We were able to get an interview with Mr. Greg Tanbara, a Health Promotion Coordinator for the Tacoma-Pierce County Health Department. Mr. Tanbara analyzes the environmental damage caused by the Asarco Plant destruction and connects with members of the community that have been affected. He told us that the state has allocated 180 million dollars to revitalizing the soil in the 1000 square mile radius most heavily contaminated by the plant’s destruction. As of today, approximately 90 million dollars have been spent on the area’s recovery, but only a small fraction of potentially dangerous areas have been tested. With the information gained from meeting with Mr. Tanbara, we learned how crucial it was for our project to be both easily useable and affordable for homeowners. The creation of an at-home test for arsenic will allow citizens to test the soil and water themselves, before requiring assistance from the health department.
  
 +
                            </p>
  
 +
  </div>
 +
</div>
 +
</div>
  
−
<p>For more information, please see the <a href="https://2018.igem.org/Human_Practices">Human Practices Hub</a>. There you will find:</p>
+
<div class="some-padding"></div>
−
+
<div class="some-padding"></div>
−
<ul>
+
−
<li> an <a href="https://2018.igem.org/Human_Practices/Introduction">introduction</a> to Human Practices at iGEM </li>
+
−
<li>tips on <a href="https://2018.igem.org/Human_Practices/How_to_Succeed">how to succeed</a> including explanations of judging criteria and advice about how to conduct and document your Human Practices work</li>
+
−
<li>descriptions of <a href="https://2018.igem.org/Human_Practices/Examples">exemplary work</a> to inspire you</li>
+
−
<li>links to helpful <a href="https://2018.igem.org/Human_Practices/Resources">resources</a></li>
+
−
<li>And more! </li>
+
−
</ul>
+
−
+
  
 +
<div class="panel-group" id="accordion" role="tablist" aria-multiselectable="true">
 +
            <div class="panel panel-default">
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 +
<div>
 +
                    <div class="col-md-11">Meeting with David Lloyd</div><div class="col-md-1"><i class="fa fa-arrow-down fa-10" aria-hidden="true"></i></div>
 +
</div>
 +
                    </a>
 +
                    </h4>
  
 +
                </div>
 +
                <div id="P3-collapse" class="panel-collapse collapse" role="tabpanel" aria-labelledby="P3">
 +
                    <div class="panel-body">
 +
<!--
 +
<p>
 +
<b>Materials:</b><br>
 +
2x Phusion Mastermix<br>
 +
10 µM forward primer<br>
 +
10 µM forward primer<br>
 +
PCR tube<br>
 +
Sterile water<br>
 +
Plasmid DNA<br>
 +
<br>
 +
<b>Methods:</b><br>
 +
For a 25 µL reaction<br>
 +
<ol style="font-size:16px;">
 +
<li>In a PCR tube on ice, combine  1-10 ng of plasmid DNA, 1.25 µL of 10 µM forward primer, 1.25 µL of 10 µM reverse primer to a PCR tube on ice, 12.5 µL of 2x Phusion Mastermix, and sterile water up to 25 µL.
 +
<br><i>Note: It is important to add Phusion Master Mix last in order to prevent primer degradation
 +
caused by the 3 ́→ 5 ́ exonuclease activity</i></li>
 +
<li>Gently mix the reaction</li>
 +
<li>If necessary, collect the liquid to the bottom of the PCR tube by spinning briefly</li>
 +
<li>Transfer the PCR tube from ice to a PCR machine to begin thermocycling</li>
 +
</ol>
  
 +
<p><br>For a 50 µL reaction<br></p>
 +
<ol style="font-size:16px;">
 +
<li>In a PCR tube on ice, combine  1-10 ng of plasmid DNA, 2.50 µL of 10 µM forward primer, 2.50 µL of 10 µM reverse primer to a PCR tube on ice, 25 µL of 2x Phusion Mastermix, and sterile water up to 50 µL.
 +
<br><i>Note: It is important to add Phusion Master Mix last in order to prevent primer degradation
 +
caused by the 3 ́→ 5 ́ exonuclease activity</i></li>
 +
<li>Gently mix the reaction</li>
 +
<li>If necessary, collect the liquid to the bottom of the PCR tube by spinning briefly</li>
 +
<li>Transfer the PCR tube from ice to a PCR machine preheated to 98°C to begin thermocycling</li>
 +
</ol>
 +
<p><br><b>Thermocycling</b><br>
 +
The PCR machine should be set to run the following steps: </p>
 +
<table class="table table-bordered table-striped">
 +
<thead>
 +
        <tr>
 +
            <th>Step</th>
 +
            <th>Temperature (°C)</th>
 +
            <th>Time</th>
 +
         
 +
        </tr>
 +
    </thead>
 +
    <tbody>
 +
        <tr>
 +
            <td>Initial denaturation</td>
 +
            <td>98</td>
 +
            <td>30 seconds</td>
 +
        </tr>
 +
        <tr>
 +
            <td>25-35 cycles</td>
 +
            <td>98 (denaturation)<br>
 +
                45-72 (annealing) <a href="#Note1">see Note 1</a><br>
 +
                72 (extension)</td>
 +
            <td>5-10 seconds <br>
 +
                10-30 seconds<br>
 +
                15-30 seconds per kb</td>
 +
        </tr>
 +
        <tr>
 +
            <td>Final extension</td>
 +
            <td>72</td>
 +
            <td>2-5 minutes</td>
 +
        </tr>
 +
        <tr>
 +
            <td>Hold</td>
 +
            <td>4</td>
 +
            <td>Indefinitely</td>
 +
        </tr>
  
−
<p>On this page, your team should document all of your Human Practices work and activities. You should write about the Human Practices topics you considered in your project, document any activities you conducted to explore these topics (such as engaging with experts and stakeholders), describe why you took a particular approach (including referencing any work you built upon), and explain if and how you integrated takeaways from your Human Practices work back into your project purpose, design and/or execution. </p>
+
    </tbody>
−
+
</table>
−
<p>If your team has gone above and beyond in work related to safety, then you should document this work on your Safety wiki page and provide a description and link on this page. If your team has developed education and public engagement efforts that go beyond a focus on your particular project, and for which would like to nominate your team for the Best Education and Public Engagement Special Prize, you should document this work on your Education and Education wiki page and provide a description and link here. </p>
+
 
 +
<p id="Note1">Note 1: Use the NEB Tm calculator should be used to determine the annealing temperature when using Phusion: <a target="_blank" href="http://tmcalculator.neb.com/#!/">http://tmcalculator.neb.com/#!/</a></p>
 +
 
 +
 
 +
</p>
 +
 
 +
  </div>
 +
</div>
 
</div>
 
</div>
−
 
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<p>The iGEM judges will review this page to assess whether you have met the Silver and/or Gold medal requirements based on the Integrated Human Practices criteria listed below. If you nominate your team for the <a href="https://2018.igem.org/Judging/Awards">Best Integrated Human Practices Special Prize</a> by filling out the corresponding field in the <a href="https://2018.igem.org/Judging/Judging_Form">judging form</a>, the judges will also review this page to consider your team for that prize.  
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                    <div class="col-md-11">Launch Party</div><div class="col-md-1"><i class="fa fa-arrow-down fa-10" aria-hidden="true"></i></div>
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</div>                   
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                    </a>
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                    <div class="panel-body">
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<p>
 +
<b>Materials:</b><br>
 +
Sterile Water<br>
 +
25 µL RedTaq mastermix<br>
 +
1 E. coli colony<br>
 +
2.5 µL of 10 µM forward primer<br>
 +
2.5 µL of 10 µM reverse primer<br>
 +
<br>
 +
<b>Methods:</b><br>
 +
<ol style="font-size:16px;">
 +
<li>Add a single colony of cells to 50 µL of water. Incubate at 95C for a minute to lyse the cells.</li>
 +
<li>Combine 1 µL cell lysate, 25 µL RedTaq mastermix, 2.5 µL of 10 µM forward primer, 2.5 µL of 10 µM reverse primer, and sterile water up to 50 µL. <br>
 +
<i>Note: It is important to add RedTaq Master Mix last in order to prevent primer
 +
degradation caused by the 3 ́→ 5 ́ exonuclease activity</i></li>
 +
<li>Incubate in the thermocycler -  Taq has a lower optimum temperature than Phusion.</li>
 +
 
 +
</ol>
 +
 
 +
<p><br><b>Thermocycling</b><br>
 +
The PCR machine should be set to run the following steps: </p>
 +
<table class="table table-bordered table-striped">
 +
<thead>
 +
        <tr>
 +
            <th>Step</th>
 +
            <th>Temperature (°C)</th>
 +
            <th>Time</th>
 +
         
 +
        </tr>
 +
    </thead>
 +
    <tbody>
 +
        <tr>
 +
            <td>Initial denaturation</td>
 +
            <td>98</td>
 +
            <td>30 seconds</td>
 +
        </tr>
 +
        <tr>
 +
            <td>25-35 cycles</td>
 +
            <td>98 (denaturation)<br>
 +
                45-72 (annealing) <a href="#Note1">see Note 1</a><br>
 +
                68 (extension)</td>
 +
            <td>5-10 seconds <br>
 +
                10-30 seconds<br>
 +
                15-30 seconds per kb</td>
 +
        </tr>
 +
        <tr>
 +
            <td>Final extension</td>
 +
            <td>72</td>
 +
            <td>5-10 minutes</td>
 +
        </tr>
 +
        <tr>
 +
            <td>Hold</td>
 +
            <td>4</td>
 +
            <td>Indefinitely</td>
 +
        </tr>
 +
 
 +
    </tbody>
 +
</table>
 +
<p>Note: If loading on a gel, the RedTaq mix contains loading dye, so don’t add anything else.</p>  
 +
-->
 +
<p>
 +
Our team was lucky enough to get a meeting with David Lloyd, CEO and Co-Founder of FREDsense Technologies. FREDsense is a Canadian startup based on the 2013 and 2012 Calgary iGEM projects. The focus of the company is to create easily usable field tests for environmental toxins. FRED (Functional Robust Electrochemical Detector) made his first appearance in 2012 alongside OSCAR (Optimized System for Carboxylic Acid Remediation). While OSCAR, the “destroy component” of the project, has not been developed since, FRED (renamed Field-Ready Electrochemical Detector) has made incredible strides since then. Today there are four FRED systems available for pre-order or trial on the FREDsense website, as well as the option of custom sensor development to fit your needs. As an iGEM veteran, current member of the iGEM Human Practices Alumni Committee, and experienced biosensor developer, Mr. Lloyd had welcome advice for us. His guidance helped us develop a plan for what our sensor needed, both ethically, and entrepreneurially.                                         
 +
                        </p>
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                    <div class="col-md-11">Meeting with Jeff Nivala</div><div class="col-md-1"><i class="fa fa-arrow-down fa-10" aria-hidden="true"></i></div>
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</div>                  
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                    </a>
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                    <div class="panel-body">
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<p>
 +
Earlier in the iGEM season, we had a morning interview and lecture with Dr. Jeff Nivala, Sr. Research Scientist and P.I. at University of Washington, as well as a recent addition to the Forbes “30 under 30” list. Dr. Nivala is most known for his research on the storage and transmission of information through molecular barcodes made of DNA and proteins. He has also been an integral part of the development of nanopore sensing technology as a method of protein analysis, which he believes to be the first step in decoding the vast complexities of whole proteomes. After recovering from our initial starstruck reactions, we were thrilled to hear his insight on our project!
 +
 
 
</p>
 
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<b>Materials:</b><br>
 +
Microcentrifuge tube<br>
 +
Media of choice<br>
 +
Overnight Culture of Cells<br>
 +
96 well plate<br>
 +
Plate reader<br>
 +
<br>
 +
<b>Methods:</b><br>
 +
<ol style="font-size:16px;">
 +
<li>Dilute overnight cultures to 0.1 OD. <b>Don’t forget the media control.</b></li>
 +
<li>Seed your cells in the 96 well plate at 100μL per well.</li>
 +
<li>Make reaction up to 20 µL using sterile water</li>
 +
<li>Set the plate reader for 600OD and run it for 12 hours for bacteria or 24 hours plus for yeast
 +
</li>
 +
</ol>
  
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<h3>Silver Medal Criterion #3</h3>
+
</div>
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<p>Convince the judges you have thought carefully and creatively about whether your work is responsible and good for the world. Document how you have investigated these issues and engaged with your relevant communities, why you chose this approach, and what you have learned. Please note that surveys will not fulfill this criteria unless you follow scientifically valid methods. </p>
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<p>Expand on your silver medal activity by demonstrating how you have integrated the investigated issues into the purpose, design and/or execution of your project. Document how your project has changed based upon your human practices work.
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                    <div class="col-md-11">Cell Sorting for Co-Culture Characterisation</div><div class="col-md-1"><i class="fa fa-arrow-down fa-10" aria-hidden="true"></i></div>
 
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                    </a>
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<p>
 +
<b>Materials:</b><br>
 +
Microcentrifuge tube<br>
 +
Media of choice<br>
 +
Overnight Culture of Cells<br>
 +
96 well plate<br>
 +
Flow Cytometer<br>
 +
PBS<br>
 +
<br>
 +
<b>Methods:</b><br>
 +
<ol style="font-size:16px;">
 +
<li>Use Flowjo to predefine the gates for the Flow cytometer. This allows the cells within the co culture to be counted based on size.</li>
 +
<li>Dilute the Cultures to 1 OD (don’t forget the media control) and culture together at desired inoculation ratio</li>
 +
<li>Set up samples in triplicate form on a 96 well plate. Dilute 10 fold and 100 fold in PBS.</li>
 +
<li>Incubate Cells and take samples at desired time points to obtain growth curves</li>
 +
</ol>
  
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<p>
 +
<b>Materials:</b><br>
 +
Replica plate or glycerol stock of cells<br>
 +
Appropriate antibiotic<br>
 +
LB broth<br>
 +
96 well plate<br>
 +
AHLs (at concentrations of 10pM, 100pM, 1nM, 10nM, 100nM, 1µM, 10µM and 100µM)<br>
 +
<br>
 +
<b>Methods:</b><br>
 +
<ol style="font-size:16px;">
 +
<li>Grow overnight culture of cells, either from replica plate or glycerol stock (see overnight culture protocol)
 +
</li>
 +
<li>In the morning, add 100 µL of the cell suspension to 10 mL LB and 10µL of antibiotic</li>
 +
<li>Grow cells in incubator for 3-4 hours at 200rpm and 37°C</li>
 +
<li>Blank spectrophotometer with 1mL LB and measure OD of cells (dilute 10x in LB)</li>
 +
<li>Make up suspension of cells at OD 0.05</li>
 +
<li>Add 100µL of cells to each well in the 96-well plate using the block-filler machine</li>
 +
<li>Using the multi-channel pipette add 2µL of intended AHL to each well, add samples in triplicate form
 +
</li>
 +
<li>Using the multi-channel pipette add 2µL of intended AHL to each well, add samples in triplicate form
 +
</li>Run on plate-reader for 12 hours, with fluorescence and absorbance measurements taken every 15 minutes (set plate reader to 37°C and 500rpm shaking)
 +
</li>
 +
</ol>
  
−
<p>To compete for the Best Integrated Human Practices prize, please describe your work on this page and also fill out the description on the judging form. </p>
+
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 +
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<p>How does your project affect society and how does society influence the direction of your project? How might ethical considerations and stakeholder input guide your project purpose and design and the experiments you conduct in the lab? How does this feedback enter into the process of your work all through the iGEM competition? Document a thoughtful and creative approach to exploring these questions and how your project evolved in the process to compete for this award!</p>
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<p>
 +
<b>PCR purification</b> was performed according to the QIAquick PCR Purification Kit<br>
 +
<b>Gel extraction</b> was performed according to the QIAquick Gel Extraction Kit<br>
 +
<b>Minipreps</b> were carried out according to the QIAprep Spin Miniprep Kit<br>
 +
 +
 +
  </div>
 
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Integrated Human Practices

As our team’s goal was to create a product for the community, integrating human practices was vitally important.
Click the titles below to see more!

Sue’s Tech Kitchen, the creation of Randi Zuckerberg, is a nationwide event dedicated to getting people excited about science through food. It’s known for its’ 3D printed smores and miracle berries, but it also serves as an opportunity for local groups to share their projects and visions with the community. We held a booth at the event in Tacoma, and had a blast telling our community all about our project. We created a mockup of our project with pH strips to represent the sensor, and solutions of a known acidity to represent water samples from around Puget Sound. Kids loved the idea of putting what looked like paper, into what looked like water, and seeing the paper change colors before their eyes. Adults were more excited about the idea of getting to test their own water and get accurate results that were easy to read. On a whiteboard, we set up a magnetic puzzle of our plasmid parts. Guests got to put together the puzzle as fast as they could, and the best time of each day won a cool water bottle. We met some incredible people, including a woman who helped create the map of arsenic contamination caused by the smelter. We had many people ask to buy test strips, but since the sensor isn’t complete, they settled for signing up for the email list. The event gave us great insight on what matters to the community, and we even got ideas on how to expand the sensor in the future!

To continue familiarizing ourselves with problems caused by arsenic contamination in the environment, we turned to our local health department. We were able to get an interview with Mr. Greg Tanbara, a Health Promotion Coordinator for the Tacoma-Pierce County Health Department. Mr. Tanbara analyzes the environmental damage caused by the Asarco Plant destruction and connects with members of the community that have been affected. He told us that the state has allocated 180 million dollars to revitalizing the soil in the 1000 square mile radius most heavily contaminated by the plant’s destruction. As of today, approximately 90 million dollars have been spent on the area’s recovery, but only a small fraction of potentially dangerous areas have been tested. With the information gained from meeting with Mr. Tanbara, we learned how crucial it was for our project to be both easily useable and affordable for homeowners. The creation of an at-home test for arsenic will allow citizens to test the soil and water themselves, before requiring assistance from the health department.

Our team was lucky enough to get a meeting with David Lloyd, CEO and Co-Founder of FREDsense Technologies. FREDsense is a Canadian startup based on the 2013 and 2012 Calgary iGEM projects. The focus of the company is to create easily usable field tests for environmental toxins. FRED (Functional Robust Electrochemical Detector) made his first appearance in 2012 alongside OSCAR (Optimized System for Carboxylic Acid Remediation). While OSCAR, the “destroy component” of the project, has not been developed since, FRED (renamed Field-Ready Electrochemical Detector) has made incredible strides since then. Today there are four FRED systems available for pre-order or trial on the FREDsense website, as well as the option of custom sensor development to fit your needs. As an iGEM veteran, current member of the iGEM Human Practices Alumni Committee, and experienced biosensor developer, Mr. Lloyd had welcome advice for us. His guidance helped us develop a plan for what our sensor needed, both ethically, and entrepreneurially.

Earlier in the iGEM season, we had a morning interview and lecture with Dr. Jeff Nivala, Sr. Research Scientist and P.I. at University of Washington, as well as a recent addition to the Forbes “30 under 30” list. Dr. Nivala is most known for his research on the storage and transmission of information through molecular barcodes made of DNA and proteins. He has also been an integral part of the development of nanopore sensing technology as a method of protein analysis, which he believes to be the first step in decoding the vast complexities of whole proteomes. After recovering from our initial starstruck reactions, we were thrilled to hear his insight on our project!

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