Team:UC Davis/Notebook/Protocols

From 2012.igem.org

(Difference between revisions)
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                    <li class='selected'        ><a href="https://2012.igem.org/Team:UC_Davis">Page</a></li>
                    <li class='selected'        ><a href="https://2012.igem.org/Team:UC_Davis">Page</a></li>
-
                 <li class='new'><a href="https://2012.igem.org/wiki/index.php?title=Talk:Team:UC_Davis&amp;action=edit&amp;redlink=1">Discussion</a></li>
+
                 <li class='new'><a href="https://2012.igem.org/wiki/index.php?title=Talk:Team:UC_Davis/Notebook/Protocols&amp;action=edit&amp;redlink=1">Discussion</a></li>
-
               <li><a href="https://2012.igem.org/wiki/index.php?title=Team:UC_Davis/Notebook/Protocols&action=edit">Edit</a></li>
+
               <li><a href="https://2012.igem.org/wiki/index.php?title=Team:UC_Davis&action=edit">Edit</a></li>
                           <li><a href="https://2012.igem.org/wiki/index.php?title=Team:UC_Davis&amp;action=history">History</a></li>
                           <li><a href="https://2012.igem.org/wiki/index.php?title=Team:UC_Davis&amp;action=history">History</a></li>
                 <li><a href="https://2012.igem.org/Special:MovePage/Team:UC_Davis">Move</a></li>
                 <li><a href="https://2012.igem.org/Special:MovePage/Team:UC_Davis">Move</a></li>
Line 372: Line 985:
         <li class="selected"><a href="https://2012.igem.org/Team:UC_Davis/Notebook" title="Notebook">Notebook</a>
         <li class="selected"><a href="https://2012.igem.org/Team:UC_Davis/Notebook" title="Notebook">Notebook</a>
           <ul>
           <ul>
-
             <li ><a href="https://2012.igem.org/Team:UC_Davis/Notebook/Protocols">Protocols</a></li>
+
             <li ><a href="https://2012.igem.org/Team:UC_Davis/Notebook_Overview ">Notebook Overview</a></li>
-
             <li ><a href="./Notebook.htm ">Week 1</a></li>
+
             <li ><a href="https://2012.igem.org/Team:UC_Davis/Notebook/Protocols ">Protocols</a></li>
-
             <li ><a href="./Notebook.htm ">Week 2</a></li>
+
             <li ><a href="https://2012.igem.org/Team:UC_Davis/Notebook">Notebook</a></li>
-
            <li ><a href="./Notebook.htm ">Week 3</a></li>
+
           </ul>
           </ul>
         </li>
         </li>
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         <li ><a href="https://2012.igem.org/Team:UC_Davis/Safety" title="Safety">Safety</a></li>
         <li ><a href="https://2012.igem.org/Team:UC_Davis/Safety" title="Safety">Safety</a></li>
-
         <li ><a href="https://2012.igem.org/Team:UC_Davis/Project" title="Project">Project</a></li>
+
         <li ><a href="https://2012.igem.org/Team:UC_Davis/Project" title="Project">Project</a>
 +
          <ul>
 +
            <li ><a href="https://2012.igem.org/Team:UC_Davis/Project">Project Overview</a></li>
 +
            <li ><a href="https://2012.igem.org/Team:UC_Davis/Project/Strain">Modular Engineering</a></li>
 +
            <li ><a href="https://2012.igem.org/Team:UC_Davis/Project/Catalyst">Chassis Engineering</a></li>
 +
            <li ><a href="https://2012.igem.org/Team:UC_Davis/Project/Protein_Engineering">Protein Engineering</a></li>
 +
          </ul>
 +
        </li>
         <li ><a href="https://2012.igem.org/Team:UC_Davis/Team" title="Team">Team</a></li>
         <li ><a href="https://2012.igem.org/Team:UC_Davis/Team" title="Team">Team</a></li>
         <li ><a href="https://2012.igem.org/Team:UC_Davis" title="Home">Home</a></li>
         <li ><a href="https://2012.igem.org/Team:UC_Davis" title="Home">Home</a></li>
     </ul>
     </ul>
   </div>
   </div>
 +
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 +
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 +
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 +
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 +
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 +
<div id="tweets">
 +
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 +
<h2>UCDavis iGEM Tweets</h2>
 +
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 +
</div>
-
<br>
+
<div id="sponsorbox" class="sponsorfloat">
-
  <div class="floatbox3">
+
<center>
-
<h1> Protocols </h1>
+
<h2>Our Sponsors</h2>
-
<article>
+
<a href="http://www.novozymes.com/en/Pages/default.aspx" target="_blank"><img src="https://static.igem.org/mediawiki/2011/2/21/UCD_Novozymes-logo.jpg" width="200"></a>
-
These are our protocols.
+
</center>
-
</article>
+
-
  </div>
+
-
<br>
+
<center>
-
  <div class="floatbox3">
+
<a href="http://engineering.ucdavis.edu/" target="_blank"><img src="https://static.igem.org/mediawiki/2011/f/f6/UCD_CoE.png" width="200" height="40"></a>
-
<article>
+
</center>
-
<hp> Registry Part Distribution Rehydration</hp>
+
 
-
<br>•Add 20 µL sterile H2O to desired well in distribution plate.
+
<center>
 +
<a href="http://biosci.ucdavis.edu/index_js.html" target="_blank"><img src="https://static.igem.org/mediawiki/2011/b/b1/UCD_biosci_sponsor.jpg" width="200" height="90"></a>
 +
</center>
 +
 
 +
<center>
 +
<a href="http://www.genomecenter.ucdavis.edu/" target="_blank"><img src="https://static.igem.org/mediawiki/2011/1/1b/UCD_Genome_center_sponsor.jpg" width="200" height="60"></a>
 +
</center>
 +
 
 +
<center>
 +
<a href="http://www.fishersci.com" target="_blank"><img src="https://static.igem.org/mediawiki/2011/a/a4/UCD_Fisher_Logo.gif" width="200"></a>
 +
</center>
 +
 
 +
<center>
 +
<a href="http://www.bme.ucdavis.edu/" target="_blank"><img src="https://static.igem.org/mediawiki/2011/4/40/UCD_BME_logo_minimal_copy.png" width="200 height="70"></a>
 +
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 +
 
 +
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 +
</div>
 +
 
 +
<div id="myleftbox">
 +
 
 +
<div id="myleftbox"  class="smallbox">
 +
<h1>Protocols</h1>
 +
<!-- accordion starts here -->
 +
<div id="firstpane" class="menu_list">
 +
 
 +
  <p class="menu_head"> Registry Part Distribution Rehydration </p>
 +
    <div class="menu_body">
 +
•Add 20 µL sterile H2O to desired well in distribution plate.
<br>•Incubate at room temperature for ~10 minutes.
<br>•Incubate at room temperature for ~10 minutes.
<br>•Transfer resuspension to microcentrifuge tube.
<br>•Transfer resuspension to microcentrifuge tube.
-
</article></div>
+
    </div>
-
<br><div class="floatbox3">
+
  <p class="menu_head"> Transformations</p>
-
<article>
+
    <div class="menu_body">
-
<hp>Transformations</hp>
+
<p>Materials</p>
<p>Materials</p>
•Competent Cells
•Competent Cells
Line 428: Line 1,140:
<br>•Place 200 µL of the transformed cells on plates containing LB and the appropriate antibiotic.
<br>•Place 200 µL of the transformed cells on plates containing LB and the appropriate antibiotic.
<br>•Incubate overnight at 37 °C.
<br>•Incubate overnight at 37 °C.
-
</article>
+
    </div>
-
  </div>
+
-
<br><div class="floatbox3">
+
  <p class="menu_head"> Restriction Enzyme Double Digest</p>
-
<article>
+
    <div class="menu_body">
-
<hp>Restriction Enzyme Double Digest</hp>
+
<p>Materials</p>
<p>Materials</p>
•22 µL dH20
•22 µL dH20
Line 446: Line 1,156:
<br>•If not purifying or running on a gel immediately, increase to 80 C for 20 minutes to kill enzymes (some enzymes need only a 65 C heatkill, check enzyme).
<br>•If not purifying or running on a gel immediately, increase to 80 C for 20 minutes to kill enzymes (some enzymes need only a 65 C heatkill, check enzyme).
<br>•Run on a gel and extract product.
<br>•Run on a gel and extract product.
-
</article></div>
+
    </div>
-
<br><div class="floatbox3">
+
  <p class="menu_head"> PCR (sequence dependent)</p>
-
<article>
+
    <div class="menu_body">
-
<hp>PCR (sequence dependent)</hp>
+
<p>Materials</p>
<p>Materials</p>
•10 uL Q Solution
•10 uL Q Solution
Line 464: Line 1,173:
•Mix reactants into PCR tubes. A large mix can be made and then aliquoted into PCR tubes if desired.
•Mix reactants into PCR tubes. A large mix can be made and then aliquoted into PCR tubes if desired.
<br>•Run in thermal cycler.
<br>•Run in thermal cycler.
-
</article></div>
+
  </div>
-
<br><div class="floatbox3">
+
  <p class="menu_head">PCR SOEing (Polymerase Chain Reaction Splicing by Overlapping Extension)</p>
-
<article>
+
    <div class="menu_body">
-
<hp>PCR SOEing (Polymerase Chain Reaction Splicing by Overlapping Extension) </hp>
+
<p>Materials</p>
<p>Materials</p>
•10 µL Q Solution
•10 µL Q Solution
Line 483: Line 1,191:
<br>•Run in thermal cycler.
<br>•Run in thermal cycler.
<br>•Continue PCR SOEing of parts until completed.
<br>•Continue PCR SOEing of parts until completed.
-
</article></div>
+
    </div>
-
</article></div>
+
-
<br><div class="floatbox3">
+
  <p class="menu_head"> Ligation</p>
-
<article>
+
    <div class="menu_body">
-
<hp>Ligation</hp>
+
<p>Materials</p>
<p>Materials</p>
•Digested Vector
•Digested Vector
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<p>Procedure</p>
<p>Procedure</p>
•Mix these materials in the amounts determined by the reaction volume calculator <a href="https://static.igem.org/mediawiki/2011/7/73/UC_Davis_Reaction_Volume_Calculator.xls">here</a>.
•Mix these materials in the amounts determined by the reaction volume calculator <a href="https://static.igem.org/mediawiki/2011/7/73/UC_Davis_Reaction_Volume_Calculator.xls">here</a>.
-
</article></div>
+
    </div>
-
<br><div class="floatbox3">
+
  <p class="menu_head"> Ethylene Glycol Media</p>
-
<article>
+
    <div class="menu_body">
-
<hp>Ethylene Glycol Media</hp>
+
<p>Materials</p>
<p>Materials</p>
•34 mM NaH<sub>2</sub>PO<sub>4</sub>
•34 mM NaH<sub>2</sub>PO<sub>4</sub>
Line 520: Line 1,225:
<br>•Mix in the ethylene glycol in a fume hood.  
<br>•Mix in the ethylene glycol in a fume hood.  
<br>•If making solid media, also mix in agar.  
<br>•If making solid media, also mix in agar.  
-
<br>•Autoclave on an appropriate cycle.  
+
<br>•Autoclave on an appropriate cycle.    
-
</article></div>
+
    </div>
-
<br><div class="floatbox3">
+
  <p class="menu_head">Ethylene Glycol Toxicity Assay</p>
-
<article>
+
    <div class="menu_body">
-
<hp>Ethylene Glycol Toxicity Assay</hp>
+
<p>Materials</p>
<p>Materials</p>
• Luria Broth Media
• Luria Broth Media
Line 531: Line 1,235:
<br>• Tecan M200 Pro
<br>• Tecan M200 Pro
<br>• E. <I>coli</I>
<br>• E. <I>coli</I>
-
 
<p>Procedure</p>
<p>Procedure</p>
• Be sure to wear a lab coat while making dilutions and aliquots, since ethylene glycol is an irritant.  
• Be sure to wear a lab coat while making dilutions and aliquots, since ethylene glycol is an irritant.  
Line 546: Line 1,249:
<br>11. Add the appropriate amount of cells to the wells for a concentration of 0.01 OD before any of the readings.  
<br>11. Add the appropriate amount of cells to the wells for a concentration of 0.01 OD before any of the readings.  
<br>12. Follow the steps of your Tecan machine appropriate to the specific bacterial strain.  
<br>12. Follow the steps of your Tecan machine appropriate to the specific bacterial strain.  
 +
    </div>
 +
</div>
 +
<!-- accordion ends here -->
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</div>
-
</article></div>
+
<!-- site map starts here -->
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<ul style="font-size:10px;list-style-image:none;list-style-type:none;float:left;display:inline;color:#000000;"  >
 +
<li style="float:left;margin:0 10px solid red"><a href="https://2012.igem.org/Team:UC_Davis"><p>Home</p><ul style="text-indent:-15px;
 +
list-style-image:none;list-style-type:none;color:#000000;"><li>Welcome </li><li>Tweets </li><li>Criteria </li><li>Sponsors </li> </ul> </a> </li>
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<li style="float:left ;margin:0 10px;"><a href="https://2012.igem.org/Team:UC_Davis/Team"><p>Team</p> <ul style="text-indent:-15px;list-style-image:none;
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list-style-type:none;color:#000000 "><li>Who we are </li><li>Students </li><li>Advisors</li></ul></a> </li>
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<li style="float:left ;margin:0 10px;"><a href="https://2012.igem.org/Team:UC_Davis/Project "><p>Project</p></a> <ul style="text-indent:-15px;list-style-image:none;list-style-type:none;color:#000000"><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Project">Project Overview</a> </li><li><a style="color:#000000 "  href="https://2012.igem.org/Team:UC_Davis/Project/Strain ">Modular Engineering</a></li><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Project/Catalyst ">Chassis Engineering</a></li><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Project/Protein_Engineering ">Protein Engineering</a></li> </ul> </li>
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<li style="float:left ;margin:0 10px"><a href="https://2012.igem.org/Team:UC_Davis/Safety "> <p>Safety</p></a> <a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Safety "> Safety</a> </li>
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<li style="float:left ;margin:0 10px;"><a href="https://2012.igem.org/Team:UC_Davis/Notebook "> <p>Notebook</p></a> <ul style="text-indent:-15px;list-style-image:none;list-style-type:none;color:#000000  "><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Notebook">Notebook Overview</a> </li><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Notebook/Protocols ">Protocols</a> </li><li><a style="color:#000000 " href="https://2012.igem.org/Team:UC_Davis/Notebook ">Notebook</a> </li> </ul> </li>
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<li style="float:left ;margin:0 10px;"><a href="https://2012.igem.org/Team:UC_Davis/Data "> <p>Data </p><ul style="text-indent:-15px;list-style-image:none;
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list-style-type:none;color:#000000"><li>Data One </li><li>Data Two </li><li>Data Three </li></ul></a> </li>
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<li style="float:left ;margin:0 10px"><a href="https://2012.igem.org/Team:UC_Davis/Attributions "> <p>Attribution </p></a><a style="color:#000000 "  href="https://2012.igem.org/Team:UC_Davis/Attributions "> Attribution</a></li>
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<li style="float:left ;margin:0 10px"><a href="https://2012.igem.org/Main_Page "> <p>iGEM </p></a><a style="color:#000000 " href="https://2012.igem.org/Main_Page " > iGEM</a></li>
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Revision as of 20:51, 4 September 2012

Team:UC Davis - 2012.igem.org

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