Team:Paris Bettencourt/Notebook/Eliminate Smell

From 2014.igem.org

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<h6>Goal: PCR agaA gBlock with oPB.001 and oPB.005</h6>
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<h6>Goal: Design plasmids that express agaA construct</h6>
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<h6>Procedure:</h6>
<h6>Procedure:</h6>
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     <p>We are going to PCR using the DreamTaq polymerase protocol the agaA gBlock with the forward primer oPB.001 and the reverse primer oPB.005. These two oligos include the BioBrick prefix and suffix. The aim is to add the prefix and suffix to the agaA sequence and amplify it.</p>
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     <p>Using Geneious, we first created the agaA construct:
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<ul>Promoter BBa_J23108 from the Anderson's promoter collection</ul>
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<ul>RBS from the RBS Calculator of Salis lab specific for E coli</ul>
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<ul>BioBrick Prefix + Scar from the iGEM Parts webpage</ul>
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<ul>agaA sequence codon optimized for E coli 12 (IDT online tool). We checked with Generious that there were no restriction sites for EcoRI, SpeI, ZbaI and PstI.</ul>
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<ul>Histidine tag: 6 Histidines in C-terminal were added</ul>
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<ul>Stop codon</ul>
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<ul>BioBrick scar + BioBrick suffix from the iGEM Parts webpage</ul>
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</p>
</br>
</br>
     <p>We will set the parameters in the machine and run the reaction overnight.</p>
     <p>We will set the parameters in the machine and run the reaction overnight.</p>

Revision as of 14:05, 19 August 2014

Paris Bettencourt 2014



Eliminate Smell

Notebook

June

June 23rd

Goal: Design plasmids that express agaA construct

Procedure:

Using Geneious, we first created the agaA construct:

    Promoter BBa_J23108 from the Anderson's promoter collection
    RBS from the RBS Calculator of Salis lab specific for E coli
    BioBrick Prefix + Scar from the iGEM Parts webpage
    agaA sequence codon optimized for E coli 12 (IDT online tool). We checked with Generious that there were no restriction sites for EcoRI, SpeI, ZbaI and PstI.
    Histidine tag: 6 Histidines in C-terminal were added
    Stop codon
    BioBrick scar + BioBrick suffix from the iGEM Parts webpage


We will set the parameters in the machine and run the reaction overnight.

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July

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August

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September

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October

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