Team:The Agency Escondido/project/rock

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<li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido">About</a></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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<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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        <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/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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        <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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<h2>
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<h2> Rock </h2>
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Project Description
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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.
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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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<h4>June 6</h4>
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<p>Yesterday's tubes were kept in the refrigerator, so they did not grow. Andrew Buss returned them to the dry bath.</p>
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<h4>June 7</h4>
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<p>The tube containing the transformation control showed excellent growth in chloramphenicol media. However, BBa_S05132 showed no growth. Andrew Buss inoculated another tube of chloramphenicol LB media with a colony from the plate.</p>
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<h4>June 10</h4>
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<p>None of Andrew Buss's BBa_S05132 tubes grew. Vansh believed that the fault was in Andrew's assembly. So, Vansh led Evan Santos and Blaise Develasco in restriction digest and ligation of <a href = "http://casp.agency-stem.org/media/G13-LP0009-21.pdf">Assembly 1 Rock Protocol</a>.</p>
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<h4>June 11</h4>
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<p>Evan Santos, Grant Jose, and Blaise Develasco prepared competent cells.</p>
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<h4>June 12</h4>
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<p>Vansh transformed BBa_S05132 and grew a chloremphenicol LB media tube.</p>
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<h4>June 13</h4>
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<p>Vansh made inoculated a chloremphenicol plate of BBa_S05132.</p>
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<h4>June 14</h4>
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<p>Vansh inoculated a new LB media + chloremphenicol tube with a single colony of BBa_S05132.</p>
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Latest revision as of 19:34, 18 June 2013