Difference between revisions of "Team:Georgia State/Collaborations"

 
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                     <div class="logo">
 
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                         <h1>Collaborations</h1>
 
                         <h1>Collaborations</h1>
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                                    <li><a href="https://2017.igem.org/Team:Georgia_State"  class="active">Home</a></li>
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            <a class="dropdown-toggle" data-toggle="dropdown">Team
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                                     <li><a href="https://2017.igem.org/Team:Georgia_State/Team">Team</a></li>
 
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                                     <li><a href="https://2017.igem.org/Team:Georgia_State/Attributions">Attributions</a></li>
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        <a href="https://2017.igem.org/Team:Georgia_State/Description" class="dropdown-toggle" data-toggle="dropdown">Project
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                                          <li><a href="https://2017.igem.org/Team:Georgia_State/Results">Results</a></li>
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                                          <li><a href="https://2017.igem.org/Team:Georgia_State/InterLab">Interlab</a></li>
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                            <li><a href="https://2017.igem.org/Team:Georgia_State/Collaborations">Collaborations</a></li>
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                                <li><a href=" https://2017.igem.org/Team:Georgia_State/Description">Description</a></li>
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                                <li><a href="https://2017.igem.org/Team:Georgia_State/Results">Results</a></li>                                        
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                <a href="https://2017.igem.org/Team:Georgia_State/Parts" class="dropdown-toggle" data-toggle="dropdown">Parts
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                                <li><a href="https://2017.igem.org/Team:Georgia_State/Collaborations">Collaborations</a></li>
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                                      <li><a href="https://2017.igem.org/Team:Georgia_State/Design">Design</a></li>
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                                      <li><a href="https://2017.igem.org/Team:Georgia_State/Parts">Parts</a></li>
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                                      <li><a href="https://2017.igem.org/Team:Georgia_State/Contribution">Contribution</a></li>
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                                          <li><a href="https://2017.igem.org/Team:Georgia_State/Improve">Improvement</a></li>
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                                    <li><a href="https://2017.igem.org/Team:Georgia_State/Safety">Safety</a></li>
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                                <li><a href="https://2017.igem.org/Team:Georgia_State/Parts">Parts</a></li>
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                                    <li><a href="https://2017.igem.org/Team:Georgia_State/Notebook">Notebook</a></li>
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                                <li><a href="https://2017.igem.org/Team:Georgia_State/Design">Design</a></li>    
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                                <li><a href="https://2017.igem.org/Team:Georgia_State/Contribution">Contribution</a></li>
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      <a href="https://2017.igem.org/Team:Georgia_State/Engagement" class="dropdown-toggle" data-toggle="dropdown">Human Practices
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                                <li><a href="https://2017.igem.org/Team:Georgia_State/Improve">Improvement</a></li>
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                            <li><a href="https://2017.igem.org/Team:Georgia_State/HP/Silver">Silver</a></li>
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                        <li><a href="https://2017.igem.org/Team:Georgia_State/HP/Gold_Integrated">Integrated and Gold</a></li>
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        <li><a href="https://2017.igem.org/Team:Georgia_State/Safety">Safety</a></li>
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                            <li><a href="https://2017.igem.org/Team:Georgia_State/Engagement">Public Engagement</a></li>
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                            <li><a href="https://igem.org/2017_Judging_Form?team=Georgia_State">Judging</a></li>
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                                </ul>
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                              <li><a href="https://2017.igem.org/Team:Georgia_State/Notebook">Notebook</a></li>  
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                            </div>
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                              <li><a href="https://2017.igem.org/Team:Georgia_State/Experiments">Experiments</a></li>
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        , <li class="dropdown-submenu">
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              <a class="dropdown-toggle" data-toggle="dropdown">Human Practices
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                          <li><a href="https://2017.igem.org/Team:Georgia_State/Engagement">Public Engagement</a></li>
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                          <li><a href="https://2017.igem.org/Team:Georgia_State/HP/Silver">Silver</a></li>
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                          <li><a href="https://2017.igem.org/Team:Georgia_State/HP/Gold_Integrated">Integrated and Gold</a>
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                              <ul><a href="https://2017.igem.org/Team:Georgia_State/HP/ASL">ASL Gallery</a></ul>
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                          </li>              
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          <li><a href="https://igem.org/2017_Judging_Form?team=Georgia_State">Judging</a></li>
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             <option value="https://2017.igem.org/Team:Georgia_State">Home</option>
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             <option value="/Team:Georgia_State">Home</option>
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             <option value="https://2017.igem.org/Team:Georgia_State">Team</option>
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            <option value="/Team:Georgia_State/Team">Team</option>
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             <option value="https://2017.igem.org/Team:Georgia_State">Project</option>
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             <option value="https://2017.igem.org/Team:Georgia_State">Parts</option>
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            <option value="/Team:Georgia_State/Results">Results</option>
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             <option value="https://2017.igem.org/Team:Georgia_State">Safety</option>
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            <option value="/Team:Georgia_State/Collaborations">Collaborations</option>
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             <option value="https://2017.igem.org/Team:Georgia_State">Human Practices</option>
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             <option value="/Team:Georgia_State/Parts">Parts</option>
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             <option value="https://2017.igem.org/Team:Georgia_State">Awards</option>
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             <option value="https://2017.igem.org/Team:Georgia_State">Notebook</option>
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             <option value="/Team:Georgia_State/Engagement">Public Engagement</option>
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            <option value="/Team:Georgia_State/HP/Silver">Silver</option>
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             <option value="/Team:Georgia_State/HP/Gold_Integrated">Gold and Integrated</option>
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            <option value="/Team:Georgia_State/HP/ASL">ASL Gallery</option>
 
             <option value="https://igem.org/2017_Judging_Form?team=Georgia_State">Judging</option>
 
             <option value="https://igem.org/2017_Judging_Form?team=Georgia_State">Judging</option>
 
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<div class="featured-blocks">
 
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<div class="media-body">
 
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<h1 class="media-heading">Mid-Atlantic Meetup</h1>
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<h1 class="media-heading">UVA Meet-Up</h1>
 
<p class="last"> The University of Virginia iGEM team invited us along with William & Mary, University of Delaware, UNC-Asheville, and the University of Maryland to the Virginia Mid-Atlantic Meetup event. Each team gave a short presentation on their project. After every two presentations, there were break-out sessions. Most of the collaboration took place during the breakout sessions. Due to this opportunity, we were able to discuss and ask questions to the other teams about their projects.  The discussions helped us think about our project on an introspective level.  Through our conversations with William & Mary, we realized the importance of the rate of protein expression in a vector and its relation to our protein expression rates.  Analyzing how each group presented improved our overall presentation technique.
 
<p class="last"> The University of Virginia iGEM team invited us along with William & Mary, University of Delaware, UNC-Asheville, and the University of Maryland to the Virginia Mid-Atlantic Meetup event. Each team gave a short presentation on their project. After every two presentations, there were break-out sessions. Most of the collaboration took place during the breakout sessions. Due to this opportunity, we were able to discuss and ask questions to the other teams about their projects.  The discussions helped us think about our project on an introspective level.  Through our conversations with William & Mary, we realized the importance of the rate of protein expression in a vector and its relation to our protein expression rates.  Analyzing how each group presented improved our overall presentation technique.
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After every presentation and break out session the Community Bio Labs from Charlottesville presented and then we had one final breakout session. This break-out session was the most important because we were able to speak directly with someone who had used HCG and pregnancy tests as a detection device. The conversation made us think about the concentration levels pregnancy tests detect and how we may need to alter our detection mechanism.</p>
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After every presentation and break out session the Community Bio Labs from Charlottesville presented and then we had one final breakout session. This break-out session was the most important because we were able to speak directly with someone who had used HCG and pregnancy tests as a detection device. The conversation made us think about the concentration levels pregnancy tests detect and how we may need to alter our detection mechanism.</p>
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<div class="media-body"><img src="https://static.igem.org/mediawiki/2017/7/7b/T--Georgia_State--UVAGROUP.jpeg" class="spacing-b no-spacing-l" alt="">
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<div class="media-body"><img src="https://static.igem.org/mediawiki/2017/7/7b/T--Georgia_State--UVAGROUP.jpeg" alt="">
 
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<h1 class="media-heading">Emory</h1>
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<p class="last">Emory, this year, had a cool project that focused on an issue faced by water refinery plants. Emory found during their visit to their local WaterHub that a primary concern was to come up with an efficient way to deal with constant fluctuating orthophosphate levels. Emory, to address this problem, decided to experiment on increasing the efficiency of organisms to eat phosphate. We assisted Emory in this endeavor by working with four strains of Bacillus subtilis. Two of the strains were wild-type isolated from water donated by the Emory Waterhub, and two of the strains were commercially available Bacillus subtilis. The goal of our work was to see which strain could uptake the most phosphate.  To test the bacteria we the protocol provided by Emory, see tab labeled Emory protocol. Then we modified the protocol, see modified protocol. And, finally, because we were lucky enough to visit the Georgia Aquarium a place with an abundant amount of water and a state of the art filtration system we asked the aquarium for some of their unfiltered and filtered water to see if they had some of the same phosphate problems.</p>
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             <div class="container">
 
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                 <div class="tabs">
 
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                     <ul id="myTab" class="nav nav-tabs">
 
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                       <li class=""><a href="#ASF" data-toggle="tab"> Atlanta Science Festival</a></li>
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                       <li class=""><a href="#Emory Protocol" data-toggle="tab"> Emory Protocol</a></li>
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                       <li class=""><a href="#STEM" data-toggle="tab">Accessibility In STEM</a></li>
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                       <li class=""><a href="#Hybrid" data-toggle="tab">GSU- Emory Protocol</a></li>
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<li class=""><a href="#More" data-toggle="tab">SBC Speaker Series</a></li>
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                                         </ul>
 
                                         </ul>
 
                     <div id="myTabContent" class="tab-content">
 
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                       <div class="tab-pane fade active in" id="ASF">
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                       <div class="tab-pane fade active in" id="Emory Protocol">
 
                         <div class="row-fluid">
 
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                                 <p class="last">We were requested to come back to ASF for 2017 due to how successful our Bio Brick activity was the previous year. We provided children and parents alike with a better understanding of the goals, applications and importance of synthetic biology by utilizing the bio brick activity and the glow fish display. This year, we educated many students and parents on synthetic biology and how to create a BioBrick construct using legos. The BioBrick activity utilizes colorful lego pieces to explain to attendees how to create a biobrick. This process involves selecting a plasmid base, promote sequence, ribosomal binding site and a coding sequence. This year used glow fish to relate how fluorescent proteins are utilized in real life. Glow fish are technicallyGMO’s that are appreciated in everyday life as a result of introducing the coding sequence from jellyfish and put it into the embryo of fish to produce a fluorescent glow. We also took some time to explained previous iGEM projects and their benefits to
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                                 <p class="last">MA6b1. cells</p>
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society. Lastly, we gave out a survey to get a better understanding of how many people are aware of the properties and benefits of CBD oil.</p>
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<p class="last"> centrifuged 2500 rpm for 5 or 10 min depending on the sample</p>
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<p class="last">  LB supernatant was dumped</p>
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<p class="last"> cells resuspended in 30 mL of 1500 uM phosphate buffer</p>
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<p class="last">centrifuged at 2500 rpm for 10 min</p>
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<p class="last">phosphate buffer supernatant dumped</p>
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<p class="last"> cells resuspended in 32 mL of 1500 uM phosphate buffer</p>
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<p class="last"> 1 mL of resuspended cells put into 1.5 mL tubes labeled with the strain and time point</p>
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<p class="last">tubes centrifuged at 10k rpm for 5 mins at the specified time point</p>
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<p class="last">specified time points</p>
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        <p class="last">t0= 30 mins</p>
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        <p class="last">t2= 1 hr</p>
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        <p class="last">t3= 2.5 hrs</p>
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        <p class="last">t4= 24 hrs?</p>
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 +
<p class="last">Leave overnight in a mixer at 37 C. </p>
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<p class="last">Next day, centrifuge 96 well plate.</p>
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<p class="last">Recipe for Malachite Green:</p>
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<p class="last">20 uL of the supernatant of each sample put into a new 96-well</p>
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<p class="last">60 uL of DI water added to each well with the sample to dilute</p>
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<p class="last">20 uL of Malachite green reagent added</p>
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<p class="last">Malachite Green - </p>
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<p class="last">500ul Malachite Green</p>
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<p class="last">125ul Ammonia </p>
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<p class="last">10ul Tween </p>
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<br></br>
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                                 <img src="https://static.igem.org/mediawiki/2017/9/90/T--Georgia_State--Phosphate.png" class="spacing-b no-spacing-l" alt="">
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<h1 class="media-heading">Source</h1>
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<p class="last">Source for picture of phosphate model: https://2017.igem.org/File:T--Georgia_State--Phosphate.png</p>
 
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                       <div class="tab-pane fade" id="STEM">
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<h3>GCDHH</h3>
+
                              <p class="last">MA6b1. cells</p>
−
                                <p class="last">Over the summer the Georgia Center of the Death and Hard-of-Hearing (GCDHH) reached out to our GSU iGEM team to learn about synthetic biology and how they could be a part of the researching world. Without knowing how to approach the obstacle of communicating with one another, we reached out to our Department of Education to find out the best way to present our information to the incoming students. Putting the guidelines we received, we hosted a successful open lab day for the GCDHH students. We had interpreters signing as we were explaining the basics of our lab and the undergraduate research that iGEM has to offer. When we initially agreed to hosting the students, we had no idea how much it would impact our view of synthetic biology. We believe that the knowledge and experience we gain from being a part of iGEM should be accessible to everyone that has an interest. Furthermore, we have created our presentation to be more accessible and have gotten an opportunity to learn sign language. </p>  
+
<p class="last"> Day One Program</p>
 +
<p class="last"> Take 5 mL of LB broth and place in 50 mL falcon tubes. Take a stab from glycerol stocks of two wild-types and two commercial strains(provided by Emory), place in tube. Let shake in the incubator overnight at 37 degrees.</p>
 +
<p class="last"> centrifuged 2500 rpm for 5 or 10 min depending on the sample</p>
 +
<p class="last">  LB supernatant was dumped</p>
 +
<p class="last"> cells resuspended in 30 mL of 1500 uM phosphate buffer</p>
 +
<p class="last">centrifuged at 2500 rpm for 10 min</p>
 +
<p class="last">phosphate buffer supernatant dumped</p>
 +
<p class="last"> cells resuspended in 32 mL of 1500 uM phosphate buffer</p>
 +
<p class="last"> 1 mL of resuspended cells put into 1.5 mL tubes labeled with the strain and time point</p>
 +
<p class="last">tubes centrifuged at 10k rpm for 5 mins at the specified time point</p>
 +
<p class="last">specified time points</p>
 +
        <p class="last">t0= 30 mins</p>
 +
        <p class="last">t2= 1 hr</p>
 +
        <p class="last">t3= 2.5 hrs</p>
 +
        <p class="last">t4= 5 hrs</p>
 +
        <p class="last">t4= 7.5 hrs</p>
 +
        <p class="last">t4= 24 hrs?</p>
 +
 
 +
<p class="last">Leave overnight in a mixer at 37 C. </p>
 +
<p class="last">Next day, centrifuge 96 well plate.</p>
 +
 
 +
 
 +
<p class="last">Recipe for Malachite Green:</p>
 +
<p class="last">20 uL of the supernatant of each sample put into a new 96-well</p>
 +
<p class="last">60 uL of DI water added to each well with the sample to dilute</p>
 +
<p class="last">20 uL of Malachite green reagent added</p>
 +
 
 +
 
 +
<br></br>
 +
 
 
<br></br>
 
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<div class="media-body"><img class="rotateimg90" src="https://static.igem.org/mediawiki/2017/b/b0/T--Georgia_State--STEMcamp5.jpg"  alt="">
 
 
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<div class="span4">
 
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<div class="media">
 
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<div class="media-body"><img  class="rotateimg90" src="https://static.igem.org/mediawiki/2017/4/4c/T--Georgia_State--STEMcamp4.jpg" height="auto" width="auto" alt="">
 
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<div class="ruler-bottom"></div>
 
−
<h3> Salvation Army Boys & Girls Club</h3> 
 
−
<p>For one of our outreach activities, the members for the iGEM team visited the salvation army boys and girls club. Here we had the pleasure of meeting with the young and curious minds. The day began with meeting all these young scientists and introducing our team and the agenda for our meetup.</p><img src="https://static.igem.org/mediawiki/2017/2/29/T--Georgia_State--boysandgirlsclubgroupphotos.jpg" align="left" hspace="20" class="spacing-b" alt="">
 
−
<p>Our basic idea was to spark interest and enthusiasm into these young minds and help them learn the importance of science in our daily life and, why it is important to the community. The meetup began with the team explaining our 2017 iGEM project in a very simple and elementary format for the kids to relate. After the explanation the team gave an elaborate breakdown of how our project can be explained and be understood in a more hands on manner using Legos. The Legos were used for the sole purpose of explaining how various BioBricks are integrated into our project.</p><div class="container"><img src="https://static.igem.org/mediawiki/2017/3/37/T--Georgia_State--boysandgirlsclubplaying.jpg" align="left" hspace="20" class="spacing-b" alt=""></div><p>The activity was wrapped up with a quick Q&A session between the students and out team where they were asked about their thoughts and reflection on how the activity shaped their understanding and if they would be interested in joining the sciences in their future endeavors or not. The meeting was finally concluded with our team handing out the fruit snacks for the kinds and thanking them for their time and patience to help us work with them.</p>
 
−
<br>
 
−
<h3>NGM/W</h3><br><p>Over the summer members of our iGEM team had an opportunity to meet with a few students from Next Generation Men and Women. NGM/W is a nonprofit Organization established to provide underrepresented students with professional exposure, leadership development, community service and personal support. Through our partnership with the group, we take on the responsibility of exposing the students to a day as a synthetic biologist. On our day, the students got to run through a typical lab day. We started off the day by autoclaving trash and loading the dishwasher with dishes. Then, we went to check on our tobacco plants; this involved testing the water level, adding food, and turning on the sun lamps in the lab. Once, the plants were taken care of; the students got to run PCR samples on a gel. While the gel ran the students got to see how to purified GFP from E.coli using hydrophobic interaction chromatography, so the students got to see GFP fluorescence. They thought this was cool, so we scored some points there! This process lasted long enough for the gel to run, and the students then got to image the gel and take a picture. This was the last bit of lab work, so we cleaned up and went to lunch. The day ended with a Q&A where we addressed any questions or doubts the students may have had about college.  We also advised the students on how to apply for scholarships, and how to fill out college applications, and how to replicate these experiments in high school. Holly and Cara, also went a step further to elaborate the diverse applications of our iGEM lab by explaining how the lab has not only a scientific component where we perform critical research but also an artistic element where students design research posters of our articulated data and present them at STEM conference. Hence providing them with an overall view of the life of an undergraduate student in STEM, and the life of an iGEMer. </p> 
 
−
<img src="https://static.igem.org/mediawiki/2017/c/ca/W.jpeg" class="spacing-b no-spacing-l" alt=""> 
 
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<h3> Interactions in Biological Systems: What are they up to? </h3> 
 
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                                <img src="https://static.igem.org/mediawiki/2017/3/37/T--Georgia_State--bhammer.jpg" align="left" hspace="20" class="spacing-b" alt="">
 
−
 
−
                                <p class="last">We hosted a lecture by microbiologist Dr. Hammer. Dr. Hammer studies cell signaling in the bacterial pathogen Vibrio cholerae, and during his talk, Dr. Hammer discussed how he uses genetic engineering for his research. His lab studies microbial interactions at scales that span genes and genomes, regulatory networks, cells, populations, and communities. Harmful and beneficial bacteria are genetically encoded with regulatory networks to integrate external information that tailors gene expression to particular niches. Bacteria use chemical signals to orchestrate behaviors that facilitate both cooperation and conflict with members of the communities they inhabit. His work focuses on the waterborne pathogen Vibrio cholera, which causes the fatal diarrheal disease cholera in humans and also resides in aquatic settings in association with other animals and surfaces like crab shells and zooplankton molts composed of chitin.</p>
 
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<hr>
 
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<img src="https://static.igem.org/mediawiki/2017/a/ac/T--Georgia_State--Aeisen.jpg" class="spacing-b" align="right" hspace="20" alt="">
 
−
<h3> CRISPR, GATTACA, and the end of the world! </h3> 
 
−
<p class="last">Arri Eisen is a Professor of Pedagogy in biology and in the Graduate Institute for Liberal Arts; he is also the Teaching Coordinator for FIRST, a National Institutes of Health-supported
 
−
postdoctoral fellowship program in research and teaching. Dr. Eisen received his undergraduate
 
−
degree in 1985 in biology with honors from UNC-Chapel Hill and his PhD in Biochemistry from
 
−
UW-Seattle in 1990. In addition to being on the Center faculty, Arri Eisen is a Professor of
 
−
Pedagogy in Biology and in the Institute for Liberal Arts; he is also the Teaching Coordinator for
 
−
FIRST, a National Institutes of Health-supported postdoctoral fellowship program in research
 
−
and teaching, and a leader of the Emory Tibet Science Initiative, which has been working over
 
−
the last decade with the Dalai Lama to educate Tibetan monks and nuns in science. Dr. Eisen
 
−
received his undergraduate degree in 1985 in biology with honors from UNC-Chapel Hill and his
 
−
PhD in Biochemistry from UW-Seattle in 1990. He has been teaching at Emory since then and
 
−
joined the Center in the late 90’s where his main responsibilities now include teaching in the
 
−
Center&'s Master of Arts in Bioethics and in Emory's Master of Science in Clinical Research
 
−
programs. Dr. Eisen publishes in the peer-reviewed literature in science, science education, and
 
−
bioethics, as well as in the popular literature. His most recent book is The Enlightened Gene:
 
−
Biology, Buddhism and the Convergence that Explains the World. Dr. Eisen spoke about
 
−
CRISPR technology and the future of creating human babies without certain medical conditions
 
−
and specific preferred traits.</p>
 
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<div class="featured-blocks"> <h2><i><font style="text-transform: none;">Our Synthetic Biology Club hosted a speaker series on campus during the spring semester.</font></i></h2></div>
 
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<br>
 
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<img src="https://static.igem.org/mediawiki/2017/b/b1/T--Georgia_State--mstyczynski.jpg" class="spacing-b" align="right" hspace="20" alt="">
 
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<h3> Learn to Engineer Bacterial Biosensors! </h3>
 
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<p class="last">The primary focus of Dr. Styczynski research is the experimental and computational study of the
 
−
dynamics and regulation of metabolism, with ultimate applications in metabolic engineering,
 
−
biotechnology, and biosensors/diagnostics. He spoke of the importance of micronutrient
 
−
deficiencies and the importance of having an accessible and affordable way to measure
 
−
deficiencies. Micronutrient deficiencies are a significant healthcare concern across the globe.
 
−
Significant even in some developed nations, micronutrient deficiencies are more severe in the
 
−
developing world and locally in the wake of major disasters. These conditions, though easily
 
−
treated, remain a problem because they are often difficult to recognize and diagnose, requiring
 
−
lab tests that are prohibitively expensive in both material and human resources for those in
 
−
developing or remote areas. As obligate consumers of the same micronutrients, bacteria possess
 
−
cellular machinery to control intracellular micronutrient levels and have corresponding
 
−
regulatory mechanisms to respond to varying concentrations in their environment. His lab is
 
−
developing a novel medical test based on bacterial sensors using designed genetic circuitry to
 
−
direct existing or minimally engineered cellular machinery to trigger specific changes in color in
 
−
response to defined micronutrient levels. Such a test would be cheap, requiring no complex
 
−
equipment and minimal medical training to administer and interpret. This would obviate the
 
−
logistical problem of laboratory access and sample transport in remote and low-resource
 
−
environments, allowing on-site diagnosis of micronutrient deficiencies in the populations most at
 
−
risk.</p>
 
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<a href="https://2017.igem.org/Team:Georgia_State"><h1 style="color:#ffffff; background-color:#A9CCE3; -moz-border-radius: 15px; -webkit-border-radius: 15px; padding:15px; text-align: center; font-family: Trebuchet MS">Back to Home</h1> </a><br><br>
  
 
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Latest revision as of 03:44, 2 November 2017

"It is the long history of humankind (and animal kind, too) those who learned to collaborate and improvise most effectively have prevailed."

Charles Darwin

UVA Meet-Up

The University of Virginia iGEM team invited us along with William & Mary, University of Delaware, UNC-Asheville, and the University of Maryland to the Virginia Mid-Atlantic Meetup event. Each team gave a short presentation on their project. After every two presentations, there were break-out sessions. Most of the collaboration took place during the breakout sessions. Due to this opportunity, we were able to discuss and ask questions to the other teams about their projects. The discussions helped us think about our project on an introspective level. Through our conversations with William & Mary, we realized the importance of the rate of protein expression in a vector and its relation to our protein expression rates. Analyzing how each group presented improved our overall presentation technique. After every presentation and break out session the Community Bio Labs from Charlottesville presented and then we had one final breakout session. This break-out session was the most important because we were able to speak directly with someone who had used HCG and pregnancy tests as a detection device. The conversation made us think about the concentration levels pregnancy tests detect and how we may need to alter our detection mechanism.

Emory

Emory, this year, had a cool project that focused on an issue faced by water refinery plants. Emory found during their visit to their local WaterHub that a primary concern was to come up with an efficient way to deal with constant fluctuating orthophosphate levels. Emory, to address this problem, decided to experiment on increasing the efficiency of organisms to eat phosphate. We assisted Emory in this endeavor by working with four strains of Bacillus subtilis. Two of the strains were wild-type isolated from water donated by the Emory Waterhub, and two of the strains were commercially available Bacillus subtilis. The goal of our work was to see which strain could uptake the most phosphate. To test the bacteria we the protocol provided by Emory, see tab labeled Emory protocol. Then we modified the protocol, see modified protocol. And, finally, because we were lucky enough to visit the Georgia Aquarium a place with an abundant amount of water and a state of the art filtration system we asked the aquarium for some of their unfiltered and filtered water to see if they had some of the same phosphate problems.

MA6b1. cells

centrifuged 2500 rpm for 5 or 10 min depending on the sample

LB supernatant was dumped

cells resuspended in 30 mL of 1500 uM phosphate buffer

centrifuged at 2500 rpm for 10 min

phosphate buffer supernatant dumped

cells resuspended in 32 mL of 1500 uM phosphate buffer

1 mL of resuspended cells put into 1.5 mL tubes labeled with the strain and time point

tubes centrifuged at 10k rpm for 5 mins at the specified time point

specified time points

t0= 30 mins

t2= 1 hr

t3= 2.5 hrs

t4= 24 hrs?

Leave overnight in a mixer at 37 C.

Next day, centrifuge 96 well plate.

Recipe for Malachite Green:

20 uL of the supernatant of each sample put into a new 96-well

60 uL of DI water added to each well with the sample to dilute

20 uL of Malachite green reagent added

Malachite Green -

500ul Malachite Green

125ul Ammonia

10ul Tween



Source

Source for picture of phosphate model: https://2017.igem.org/File:T--Georgia_State--Phosphate.png

MA6b1. cells

Day One Program

Take 5 mL of LB broth and place in 50 mL falcon tubes. Take a stab from glycerol stocks of two wild-types and two commercial strains(provided by Emory), place in tube. Let shake in the incubator overnight at 37 degrees.

centrifuged 2500 rpm for 5 or 10 min depending on the sample

LB supernatant was dumped

cells resuspended in 30 mL of 1500 uM phosphate buffer

centrifuged at 2500 rpm for 10 min

phosphate buffer supernatant dumped

cells resuspended in 32 mL of 1500 uM phosphate buffer

1 mL of resuspended cells put into 1.5 mL tubes labeled with the strain and time point

tubes centrifuged at 10k rpm for 5 mins at the specified time point

specified time points

t0= 30 mins

t2= 1 hr

t3= 2.5 hrs

t4= 5 hrs

t4= 7.5 hrs

t4= 24 hrs?

Leave overnight in a mixer at 37 C.

Next day, centrifuge 96 well plate.

Recipe for Malachite Green:

20 uL of the supernatant of each sample put into a new 96-well

60 uL of DI water added to each well with the sample to dilute

20 uL of Malachite green reagent added








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