Team:Trieste/notebook

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</br>  
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We extracted the following BB from the kit: the constitutive promoter (BBa_J23100), the strong RBS (BBa_B0034), the weak RBS (BBa_B0031), the GFP (BBa_E0240) the double terminator (BBa_B0015), the toxin Tse2 (BBa_K314200).</br>
We extracted the following BB from the kit: the constitutive promoter (BBa_J23100), the strong RBS (BBa_B0034), the weak RBS (BBa_B0031), the GFP (BBa_E0240) the double terminator (BBa_B0015), the toxin Tse2 (BBa_K314200).</br>
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</br>  
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<i>E.coli</i> DH5-alpha competent cells were transformed with 2 ul of each BB extracted.</br>  
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E.coli DH5-alpha competent cells were transformed with 2 ul of each BB extracted.</br>  
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</br>  
</br>  
We learnt the safety rules and mostly of the lab techniques.  
We learnt the safety rules and mostly of the lab techniques.  
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    <div id="antibody" class="notebook_section">
    <div id="antibody" class="notebook_section">
    <h2 class="notebook_title">Antibody</h2>
    <h2 class="notebook_title">Antibody</h2>
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The first week in the lab we prepared competent E. coli strains DH5L, W3110 and HB2151 necessary for future experiments. Competent W3110 and HB2151were first transformed with p-REP 4 plasmid which codes for Lac repressor. The recombinant colonies selected on plate containing kanamycin were inoculated the and made them competent.</br></br>  
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The first week in the lab we prepared competent <i>E. coli</i> strains DH5-alpha, W3110 and HB2151 necessary for future experiments. Competent W3110 and HB2151 were first transformed with p-REP 4 plasmid which codes for Lac repressor. The recombinant colonies selected on plate containing kanamycin were inoculated the and made them competent.</br></br>  
We also digested and purified the synthetized genes from pUC57 vector.
We also digested and purified the synthetized genes from pUC57 vector.
    </div>
    </div>
    <div id="chassis" class="notebook_section">
    <div id="chassis" class="notebook_section">
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    <h2 class="notebook_title">Chassis</h2>
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    <h2 class="notebook_title">Cumate-Switch Regulation</h2>
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We received the two plasmids that we had built and ordered.
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We waited the synthetic DNA (RBS - CymR - SV5 tag and T5 promoter - Cumate operator).
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The first one contains the RBS - CymR - SV5 tag (730bp), the second one contain T5 promoter - Cumate operator (129 bp).
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    </div>
    </div>
                 </div>
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             <div id="box_right"> <!-- start box_right -->
             <div id="box_right"> <!-- start box_right -->
             <ul id="sub_menu">
             <ul id="sub_menu">
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    <li><a href="https://2012.igem.org/Team:Trieste/notebook">Week 1</a></li>
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    <li class="select" ><a href="https://2012.igem.org/Team:Trieste/notebook">Week 1</a></li>
                     <li><a href="https://2012.igem.org/Team:Trieste/notebook2">Week 2</a></li>
                     <li><a href="https://2012.igem.org/Team:Trieste/notebook2">Week 2</a></li>
                     <li><a href="https://2012.igem.org/Team:Trieste/notebook3">Week 3</a></li>
                     <li><a href="https://2012.igem.org/Team:Trieste/notebook3">Week 3</a></li>
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                     <li><a href="https://2012.igem.org/Team:Trieste/notebook11">Week 11</a></li>
                     <li><a href="https://2012.igem.org/Team:Trieste/notebook11">Week 11</a></li>
                     <li><a href="https://2012.igem.org/Team:Trieste/notebook12">Week 12</a></li>
                     <li><a href="https://2012.igem.org/Team:Trieste/notebook12">Week 12</a></li>
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                    <li><a href="https://2012.igem.org/Team:Trieste/notebooklast">Post European-Jamboree</a></li>
                 </ul>
                 </ul>
                 <img src="https://static.igem.org/mediawiki/2012/b/b0/Team_trieste.jpg" alt="Team iGEM 2012" id="igem_team" />
                 <img src="https://static.igem.org/mediawiki/2012/b/b0/Team_trieste.jpg" alt="Team iGEM 2012" id="igem_team" />
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Latest revision as of 18:05, 26 October 2012

Week 1

More

Suicide System

We prepared the antibiotics stock ( Amp, Kan, Cm, Spec ) and the DH5-alpha competent cells.

We extracted the following BB from the kit: the constitutive promoter (BBa_J23100), the strong RBS (BBa_B0034), the weak RBS (BBa_B0031), the GFP (BBa_E0240) the double terminator (BBa_B0015), the toxin Tse2 (BBa_K314200).
E.coli DH5-alpha competent cells were transformed with 2 ul of each BB extracted.

We learnt the safety rules and mostly of the lab techniques.

Antibody

The first week in the lab we prepared competent E. coli strains DH5-alpha, W3110 and HB2151 necessary for future experiments. Competent W3110 and HB2151 were first transformed with p-REP 4 plasmid which codes for Lac repressor. The recombinant colonies selected on plate containing kanamycin were inoculated the and made them competent.

We also digested and purified the synthetized genes from pUC57 vector.

Cumate-Switch Regulation

We waited the synthetic DNA (RBS - CymR - SV5 tag and T5 promoter - Cumate operator).
Università degli studi di Trieste ICGEB Illy Fondazione Cassa di Risparmio
iGEM 2012 iGEM 2012 iGEM 2012 iGEM 2012 iGEM 2012 iGEM 2012
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