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

 
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                         <h1>Collaborations</h1>
 
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                              <ul><a href="https://2017.igem.org/Team:Georgia_State/HP/ASL">ASL Gallery</a></ul>
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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">Parts</option>
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            <option value="/Team:Georgia_State/Contribution">Contribution</option>
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             <option value="/Team:Georgia_State/Improvement">Improve</option>
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             <option value="/Team:Georgia_State/Notebook">Notebook</option>
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            <option value="/Team:Georgia_State/Experiments">Experiments</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>
 
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<h1 class="media-heading">Emory</h1>
 
<h1 class="media-heading">Emory</h1>
 
<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>
 
<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>
 +
</div>
 +
            </div>
 +
            <div class="container">
 +
                <div class="tabs">
 +
                    <ul id="myTab" class="nav nav-tabs">
 +
                      <li class=""><a href="#Emory Protocol" data-toggle="tab"> Emory Protocol</a></li>
 +
                      <li class=""><a href="#Hybrid" data-toggle="tab">GSU- Emory Protocol</a></li>
  
</div>
+
                                        </ul>
 +
                    <div id="myTabContent" class="tab-content">
 +
                      <div class="tab-pane fade active in" id="Emory Protocol">
 +
                        <div class="row-fluid">
 +
                            <div class="span6 ruler-right ruler-bottom">
 +
                                <div class="block">
 +
                                <p class="last">MA6b1. cells</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= 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>
  
</div>
 
  
 +
<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>
 +
 +
<p class="last">Malachite Green - </p>
 +
<p class="last">500ul Malachite Green</p>
 +
<p class="last">125ul Ammonia </p>
 +
<p class="last">10ul Tween </p>
 +
<br></br>
 +
 +
                                </div>
 +
                            </div>
 +
                            <div class="span6">
 +
                                <div class="block no-spacing-l">
 +
                                <img src="https://static.igem.org/mediawiki/2017/9/90/T--Georgia_State--Phosphate.png" class="spacing-b no-spacing-l" alt="">
 +
<h1 class="media-heading">Source</h1>
 +
<p class="last">Source for picture of phosphate model: https://2017.igem.org/File:T--Georgia_State--Phosphate.png</p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
                      </div>
 +
                      <div class="tab-pane fade" id="Hybrid">
 +
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 +
                            <div class="span10 ruler-bottom">
 +
                                <div class="block">
 +
                              <p class="last">MA6b1. cells</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>
 +
</div>
 +
                         
 +
</div>
 
  </div>
 
  </div>
 
                         </div>
 
                         </div>
 
                           </div>
 
                           </div>
 +
<br><br><br>
 +
<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

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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