Team:The Agency Escondido/project/rock

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<ul>
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<ul id="nav">
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<li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido">About</a></li>
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<li>
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<li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido/notebook">Notebook</a></li>
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    <a href="https://2013hs.igem.org/Team:The_Agency_Escondido">About</a>
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<li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido/safety">Safety</a></li>
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    <ul>
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<li id="selectedsection">Project</a></li>
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<li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido/sandiego">San Diego</a></li>
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<li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido/ourteam">Our Team</a></li>
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<li>
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    <a href="https://2013hs.igem.org/Team:The_Agency_Escondido/notebook">Notebook</a>
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    <a href="https://2013hs.igem.org/Team:The_Agency_Escondido/safety">Safety</a>
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        <li></li>
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<li>
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    <a href="https://2013hs.igem.org/Team:The_Agency_Escondido/project"><u>Project</u></a>
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        <li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido/project">Overveiw</a></li>
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        <li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido/project/rock">Rock</a></li>
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        <li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido/project/paper">Paper</a></li>
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        <li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido/project/scissors">Scissors</a></li>
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<li>
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    <a href="https://2013hs.igem.org/Team:The_Agency_Escondido/sandiego">San Diego</a>
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    <ul>
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        <li></li>
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    </ul>
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</li>
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<li>
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    <a href="https://2013hs.igem.org/Team:The_Agency_Escondido/ourteam">Our Team</a>
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Our project seeks to create two identical triads of transparent bacteria strains with complementary hydrophobic inducer production to emulate the game of "rock, paper, scissors" (Hydrophobic proteins can move through plasma membranes). Each triad will correspond to a group: group A and group B. Each type of bacteria will produce a specific protein that will act as an inducer to one and only one other bacteria strain(rock strain, paper strain, and scissors strain). Each triad will have different reporter genes designating its champion color. When two bacteria strains are combined in an agar plate, each protein secreted by the victorious strain, when acting as an inducer, changes the color of the cells on the plate identifying the champion.
Our project seeks to create two identical triads of transparent bacteria strains with complementary hydrophobic inducer production to emulate the game of "rock, paper, scissors" (Hydrophobic proteins can move through plasma membranes). Each triad will correspond to a group: group A and group B. Each type of bacteria will produce a specific protein that will act as an inducer to one and only one other bacteria strain(rock strain, paper strain, and scissors strain). Each triad will have different reporter genes designating its champion color. When two bacteria strains are combined in an agar plate, each protein secreted by the victorious strain, when acting as an inducer, changes the color of the cells on the plate identifying the champion.
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<img height="650px" width="700px" src=https://static.igem.org/mediawiki/2013hs/a/af/The_agency_escondido_plasmid_map_2013_2.gif></img>
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<a id="rock"></a>
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<table align="center" border="0">
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    <th><h2>Contents</h2></th>
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      <td><a href="#rock"> Rock </a> </td>
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      <td><a href="#paper"> Paper </a> </td>
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      <td><a href="#scissors" > Scissors </a> </td> 
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<a id="rock"></a>
<h2> Rock </h2>
<h2> Rock </h2>
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<p>The "rock" bacteria strain will have a CFP (<a href = "http://partsregistry.org/Part:BBa_E0020">part E0020</a>) and promoter (<a href = "http://partsregistry.org/Part:BBa_R0080">part R0080</a>). It will contain the LuxR gene (<a href = "http://partsregistry.org/Part:BBa_C0062">part C0062</a>) to induce the "paper" strain.</p>
<p>The "rock" bacteria strain will have a CFP (<a href = "http://partsregistry.org/Part:BBa_E0020">part E0020</a>) and promoter (<a href = "http://partsregistry.org/Part:BBa_R0080">part R0080</a>). It will contain the LuxR gene (<a href = "http://partsregistry.org/Part:BBa_C0062">part C0062</a>) to induce the "paper" strain.</p>
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<p>Andrew Buss completed ligation of the restriction digests. While he had intended to transform immediately, Evan Santos' cell competency protocol went awry, and competent cells were not available. Transformation should take place during the next day.</p>
<p>Andrew Buss completed ligation of the restriction digests. While he had intended to transform immediately, Evan Santos' cell competency protocol went awry, and competent cells were not available. Transformation should take place during the next day.</p>
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<h4>June 4</h4>
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<p>Andrew Buss prepared a glycerol stock of pSB1C3 for long term storage from sample 11.</p>
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<p>Andrew Buss prepared competent cells using <a href="http://casp.agency-stem.org/media/G13-LP0012-30.pdf">revision 30 of the cell competency protocol by Evan Santos</a>.</p>
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<p>Andrew Buss transformed the sample of BBa_S05132 ligated the previous day (sample 28) into the previously prepared competent cells, using <a href="http://casp.agency-stem.org/media/G13-LP0014-5.pdf">revision 5 of the calcium chloride transformation protocol by Vansh Singh</a>. Additionally, 10 uL of 20 pg/uL RFP control was used as a transformation control. After 1 hour of growth in NEB SOC media, the cells were applied to chloramphenicol plates included with the 3A Assembly kit, and spread around with glass beads.</p>
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<h4>June 5</h4>
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<p>Vansh inoculated tubes with 5ml LB + chloremphenicol of the two plates from yesterday.</p>
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Revision as of 00:25, 7 June 2013