Team:Arizona State/notebook

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<tr> <td colspan="9"><a href="https://2014.igem.org/Main_Page" style="width:65px; float:right; margin-left:5px"> <img src="https://static.igem.org/mediawiki/igem.org/6/60/Igemlogo_300px.png" width="55px"></a>
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  <li><a href="https://2014.igem.org/Team:Arizona_State">home </a></li>
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  <li><a href="https://2014.igem.org/Team:Arizona_State/team">team </a>
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      <ul><li><a href="https://2014.igem.org/Team:Arizona_State/members">members </a></li>
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  <li><a href="https://2014.igem.org/Team:Arizona_State/project">project </a>
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        <li><a href="https://2014.igem.org/Team:Arizona_State/policypractices">policy and practices</a></li></ul></li>
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  <li><a href="https://2014.igem.org/Team:Arizona_State/parts">parts </a></li>
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  <li><a href="https://2014.igem.org/Team:Arizona_State/result">results </a></li>
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  <li><a href="https://2014.igem.org/Team:Arizona_State/notebook">notebook </a></li>
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  <li><a href="https://2014.igem.org/Team:Arizona_State/safety">safety </a></li>
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<a href="https://2014.igem.org/Team:Arizona_State/notebook"style="color:#ffffff">notebook </a> </td>
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<td width="90%" ><p><strong style="font-size: 34px">Safety Procedures</strong></p>
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<td width="90%" ><p><strong style="font-size: 34px">Notebook</strong></p>
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   <p>&nbsp;</p>
   <p>&nbsp;</p>
   <p><strong>Project Phases</strong> <br>
   <p><strong>Project Phases</strong> <br>
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<p>&nbsp;</p>
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     <p>June: Project planning and set up</p>
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<p>July-October: Synthesis of Fatty acid producing plasmids, such as the TesA plasmid</p>
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<p>September - October: Preparation and testing of the ethanol producing parts</p>
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<p>October: Preparation and testing of the wax esterase plasmid</p>
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   <p>&nbsp;</p>
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   <p><strong>Procedures</strong> <br>
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     Gel making/running.  
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     Gel making/running: Gels were all 1% agarose gels using safe white dye. Al were ran for 1 hr. at 95 V.
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    Enyzme digestions/ Ligations: Our team had a bit of a learning curve with digestions and ligations. We tried several different methods to ensure they would work, but the most common was the procedure found in the <a href="http://openwetware.org/wiki/Haynes:Assembly101">Haynes Lab Procedures</a>
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Enyzme digestions.
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<p>&nbsp;</p>
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  <p><strong>Resting Cell Assay</strong> <p>
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    To test the productivity of different plasmids, resting cell assays were run. All assays were run in a shaker incubator at 32 degrees C and 200 RPM. A phosphate buffered solution was used to suspend the growth of the cultures.  </p>
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<p>&nbsp;</p>
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  <p><strong>Quantification</strong> <p>
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A GC with a DB5 column was used to quantify data from all assays. For testing ethanol production, the method used had an initial temp. of 60 degrees C, and ramped by 10 degrees C until it reached 250 degrees C, which was then held for 3 minutes.
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Ligations.
 
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Testing the parts (David).
 
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  <p><strong>Daily entries from open wet ware?</strong> <br>
 
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Latest revision as of 03:08, 18 October 2014

 

 

Notebook

 

Project Phases

 

June: Project planning and set up

July-October: Synthesis of Fatty acid producing plasmids, such as the TesA plasmid

September - October: Preparation and testing of the ethanol producing parts

October: Preparation and testing of the wax esterase plasmid

 

 

Procedures
Gel making/running: Gels were all 1% agarose gels using safe white dye. Al were ran for 1 hr. at 95 V.

Enyzme digestions/ Ligations: Our team had a bit of a learning curve with digestions and ligations. We tried several different methods to ensure they would work, but the most common was the procedure found in the Haynes Lab Procedures

 

Resting Cell Assay

To test the productivity of different plasmids, resting cell assays were run. All assays were run in a shaker incubator at 32 degrees C and 200 RPM. A phosphate buffered solution was used to suspend the growth of the cultures.

 

Quantification

A GC with a DB5 column was used to quantify data from all assays. For testing ethanol production, the method used had an initial temp. of 60 degrees C, and ramped by 10 degrees C until it reached 250 degrees C, which was then held for 3 minutes.

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