Difference between revisions of "Team:CMUQ/protocols"

 
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                 <a class="dropdown-toggle" data-toggle="dropdown" href="#">PROJECT <span class="caret"> </span></a>
 
                 <a class="dropdown-toggle" data-toggle="dropdown" href="#">PROJECT <span class="caret"> </span></a>
 
                 <ul class="dropdown-menu">
 
                 <ul class="dropdown-menu">
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                            <li><a href="https://2017.igem.org/Team:CMUQ/descritpion">Description</a></li>
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  <li><a href="https://2017.igem.org/Team:CMUQ/Description">Description</a></li>
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                             <li><a href="https://2017.igem.org/Team:CMUQ/design">Design</a></li>
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                             <li><a href="https://2017.igem.org/Team:CMUQ/Design">Design</a></li>
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                             <li><a href="https://2017.igem.org/Team:CMUQ/safety">Safety</a></li>
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                             <li><a href="https://2017.igem.org/Team:CMUQ/Safety">Safety</a></li>
 
                             <li><a href="https://2017.igem.org/Team:CMUQ/Demonstrate">Demonstrate</a></li>
 
                             <li><a href="https://2017.igem.org/Team:CMUQ/Demonstrate">Demonstrate</a></li>
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                             <li><a href="https://2017.igem.org/Team:CMUQ/Model">Model</a></li>
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                             <li><a href="https://2017.igem.org/Team:CMUQ/Parts">Parts</a></li>
 
                             <li><a href="https://2017.igem.org/Team:CMUQ/Collaborations">Collaborations</a></li>
 
                             <li><a href="https://2017.igem.org/Team:CMUQ/Collaborations">Collaborations</a></li>
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                             <li><a href="https://2017.igem.org/Team:CMUQ/achievments">Achievements</a></li>
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                             <li><a href="https://2017.igem.org/Team:CMUQ/JudgingForm">Judging Form </a></li>
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                 </ul>
 
                 </ul>
 
             </li>
 
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             <li class="dropdown">
 
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                 <a class="dropdown-toggle" data-toggle="dropdown" href="#"> PRACTICES <span class="caret"> </span></a>
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                 <a class="dropdown-toggle" data-toggle="dropdown" href="#"> COMMUNITY INVOLVEMENT <span class="caret"> </span></a>
 
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                             <li><a href="https://2017.igem.org/Team:CMUQ/humanpractices">Human Practices</a></li>
 
                             <li><a href="https://2017.igem.org/Team:CMUQ/humanpractices">Human Practices</a></li>
 
                             <li><a href="https://2017.igem.org/Team:CMUQ/HP/Silver"> Silver HP </a></li>
 
                             <li><a href="https://2017.igem.org/Team:CMUQ/HP/Silver"> Silver HP </a></li>
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                            <li><a href="https://2017.igem.org/Team:CMUQ/HP/Gold_Integrated"> Gold HP </a></li>
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                            <li><a href="https://2017.igem.org/Team:CMUQ/Engagement"> Outreach</a></li>
 
                 </ul>
 
                 </ul>
 
             </li>
 
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             <li class="dropdown">
 
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                 <a class="dropdown-toggle" data-toggle="dropdown" href="#"> WET LAB <span class="caret"> </span></a>
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                 <a class="dropdown-toggle" data-toggle="dropdown" href="#"> LAB WORK <span class="caret"> </span></a>
 
                 <ul class="dropdown-menu">
 
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                             <li><a href="https://2017.igem.org/Team:CMUQ/results">Overview & Results</a></li>
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                             <li><a href="https://2017.igem.org/Team:CMUQ/notebook">Notebook</a></li>
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                             <li><a href="https://2017.igem.org/Team:CMUQ/Notebook">Notebook</a></li>
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                             <li><a href="https://2017.igem.org/Team:CMUQ/protocols">Protocols</a></li>
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                             <li><a href="https://2017.igem.org/Team:CMUQ/protocols">Procedures</a></li>
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                             <li><a href="https://2017.igem.org/Team:CMUQ/InterLab">InterLab</a></li>
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                             <li><a href="https://2017.igem.org/Team:CMUQ/InterLab">InterLab Study</a></li>
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                </ul>
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                            <li><a href="https://2017.igem.org/Team:CMUQ/Results">Results</a></li>
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            </li>
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                <a class="dropdown-toggle" data-toggle="dropdown" href="#">DRY LAB<span class="caret"> </span></a>
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                    <li><a href="https://2017.igem.org/Team:CMUQ/modelling">Modelling</a></li>
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                 </ul>
 
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                     <li><a href="https://2017.igem.org/Team:CMUQ/team">Team</a></li>
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                             <li><a href="https://2017.igem.org/Team:CMUQ/Attributions">Attributions</a></li>
 
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<div class="centered"> <h1>Protocols </h1> </div>
 
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<button type="button" class="btn btn-default btn-lg btn-block" style= "font-size: 35px;"><a href="https://2017.igem.org/Team:CMUQ/BioSensor_Lab"> BioSensor Lab<a/></button>
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<button type="button" class="btn btn-default btn-lg btn-block" style= "font-size: 35px;"><a href="https://2017.igem.org/Team:CMUQ/Dspb_Lab">DspB Lab<a/></button>
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<button type="button" class="btn btn-default btn-lg btn-block" style= "font-size: 35px;"><a href="https://2017.igem.org/Team:CMUQ/Biofilm_Formation_Lab">Biofilm Formation Lab<a/></button>
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<button type="button" class="btn btn-default btn-lg btn-block" style= "font-size: 35px;"><a href="https://2017.igem.org/Team:CMUQ/InterLab">Interlab<a/></button>
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<h3>A1: T7 salt sensor </h3>
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<h4> Introduction </h4>
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<p> The goal is to amplify T7 salt sensor, digest it along with pSB1C3 plasmid with EcoRI and PstI</p>
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<h4>Materials </h4>
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<ul>
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<li> Polymerase: Fusion Taq or Ampli tas (gold) </li>
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<li>Primers: PrefixF  and SuffixR  </li>
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<li>Nuclease free H2O </li>
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<li>5xHF PCR Buffer </li>
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<li>gBlock DNA </li>
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<li> dNTPs</li>
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<li>PCR tubes </li>
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</ul>
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<h4> Procedure </h4>
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<p style= "font-size: 20px;"> Preparation of reagents </p>
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<ol>
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<li> Resuspend gBLOCK in 20µL nuclease free dH2O</li>
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<li> Resuspend primers to make 100µM stock then dilute to make a 10µM stock</li>
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<li>Add the following reagents to a PCR tube: </li>
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<img src="https://static.igem.org/mediawiki/2017/e/ef/Table_a1.png" width = 35% position="center">
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<li>Run the PCR machine </li>
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<img src="https://static.igem.org/mediawiki/2017/2/21/A1_table2.png" width = 45% position="center">
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<li> Digest amplified sequence with EcoRI and PstI </li>
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<li> Digest pSB1C3 vector with EcoRI and PstI</li>
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<li> If you clone into a vector that expresses a fluorescent protein, other than red since you are cloning in red, then colonies that still express that fluorescent protein will be negative clones and the red or white colonies will be the ones that you want.</li>
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<li>Run on an agarose gel, cut out the bands and purify.  </li>
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<li> Send clones for sequencing  </li>
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</ol>
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<img src = "https://static.igem.org/mediawiki/2017/e/e5/Occidental_Logo..png" width = "15%;" >
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<img src = "https://static.igem.org/mediawiki/2017/f/fa/IDT_logo.cmuq.png" width = "20%;" >
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Latest revision as of 01:41, 2 November 2017

Protocols