Difference between revisions of "Team:Aalto-Helsinki/LabBook"

m (fixing lab book)
m (project titles to tables)
Line 224: Line 224:
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Plates</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Preparations <br/> Plates</h3></div></td>
 
         <td class="white arrow" >
 
         <td class="white arrow" >
 
             <p>Making Cm <br/>plates</p>
 
             <p>Making Cm <br/>plates</p>
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   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Competent Cells</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Preparations <br/> Competent Cells</h3></div></td>
 
         <td class="white arrow" >
 
         <td class="white arrow" >
 
             <p>Preparing <br/>solutions for <br/>competent cells</p>
 
             <p>Preparing <br/>solutions for <br/>competent cells</p>
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   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Competent Cells</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3></h3></div></td>
 
         <td class="white arrow" >
 
         <td class="white arrow" >
 
             <p>o/n culture <br/> of E. coli <br/> TOP10</p>
 
             <p>o/n culture <br/> of E. coli <br/> TOP10</p>
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   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Competent Cells</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3></h3></div></td>
 
         <td class="white arrow" >
 
         <td class="white arrow" >
 
             <p>o/n culture <br/> of E. coli <br/> BL21(DE3)</p>
 
             <p>o/n culture <br/> of E. coli <br/> BL21(DE3)</p>
Line 269: Line 269:
 
   </tr>
 
   </tr>
 
     <tr>
 
     <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Propagations</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Preparations <br/>Propagations</h3></div></td>
 
         <td class="white arrow" >
 
         <td class="white arrow" >
 
             <p>Transformed <br/> &nbsp;&nbsp;AH001-AH008 <br/> into TOP10 </p>
 
             <p>Transformed <br/> &nbsp;&nbsp;AH001-AH008 <br/> into TOP10 </p>
Line 309: Line 309:
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Propagations</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Preparations <br/>Propagations</h3></div></td>
 
         <td class="white arrow" >
 
         <td class="white arrow" >
 
             <p style="font-size:12px">Transformed <br/>AH007-AH009,<br/> AH005 & AH019 <br/> into TOP10</p>
 
             <p style="font-size:12px">Transformed <br/>AH007-AH009,<br/> AH005 & AH019 <br/> into TOP10</p>
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         <td class="blue arrow" >
 
         <td class="blue arrow" >
 
             <p>MiniPrep</p>
 
             <p>MiniPrep</p>
 +
        </td>
 +
  </tr>
 +
  <tr>
 +
        <td class="first rightline"><div class="heightdiv"><h3></h3></div></td>
 +
        <td class="white arrow" >
 +
            <p style="font-size:12px">o/n culture of<br/>&nbsp;AH001-AH004 <br/> & AH006</p>
 +
        </td>
 +
        <td class="blue arrow" >
 +
            <p >MiniPrep</p>
 
         </td>
 
         </td>
 
   </tr>
 
   </tr>
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   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Competent Cells</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Checking <br/>Competent Cells</h3></div></td>
 
         <td class="white arrow" >
 
         <td class="white arrow" >
 
             <p style="font-size:12px">o/n culture <br/> of BL21(DE3) <br/> transformants</p>
 
             <p style="font-size:12px">o/n culture <br/> of BL21(DE3) <br/> transformants</p>
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   </tr>
 
   </tr>
 
     <tr>
 
     <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Assembly<br/> Testing RBS's</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Testing RBS's <br/> Assembly</h3></div></td>
 
         <td class="white arrow" >
 
         <td class="white arrow" >
 
             <p style="font-size:12px">Restrict <br/>AH001-AH004 <br/> & AH006-AH008</p>
 
             <p style="font-size:12px">Restrict <br/>AH001-AH004 <br/> & AH006-AH008</p>
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             <p>Plates in rt <br/> over weekend</p>
 
             <p>Plates in rt <br/> over weekend</p>
 
         </td>   
 
         </td>   
  </tr>
 
 
  <tr>
 
        <td class="first rightline"><div class="heightdiv"><h3>Propagations</h3></div></td>
 
        <td class="white arrow" >
 
            <p style="font-size:12px">o/n culture of<br/>&nbsp;&nbsp;AH001-AH004 <br/> & AH006</p>
 
        </td>
 
        <td class="blue arrow" >
 
            <p >MiniPrep</p>
 
        </td>
 
 
   </tr>
 
   </tr>
  
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   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Assembly <br/> Testing RBS's</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Preparations <br/>Propagations</h3></div></td>
 +
        <td class="white arrow" >
 +
            <p style="font-size:13px;">Transform <br/>&nbsp;&nbsp;&nbsp; AH020 & AH021 <br/> into TOP10</p>
 +
        </td>
 +
        <td class="white arrow" >
 +
            <p>Plates in rt <br/> &nbsp;over weekend</p>
 +
        </td>
 +
  </tr>
 +
  <tr>
 +
        <td class="first rightline"><div class="heightdiv"><h3>Testing RBS's <br/> Assembly</h3></div></td>
  
 
         <td class="red arrow" >
 
         <td class="red arrow" >
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   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
<td class="first rightline"><div class="heightdiv"><h3>Assembly <br/> Testing RBS's</h3></div></td>
+
<td class="first rightline"><div class="heightdiv"><h3></h3></div></td>
  
 
         <td class="blue arrow" >
 
         <td class="blue arrow" >
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         <td class="blue arrow" >
 
         <td class="blue arrow" >
 
             <p>MiniPrep</p>
 
             <p>MiniPrep</p>
        </td>
 
  </tr>
 
  <tr>
 
        <td class="first rightline"><div class="heightdiv"><h3>Propagations</h3></div></td>
 
        <td class="white arrow" >
 
            <p style="font-size:13px;">Transform <br/>&nbsp;&nbsp;&nbsp; AH020 & AH021 <br/> into TOP10</p>
 
        </td>
 
        <td class="white arrow" >
 
            <p>Plates in rt <br/> &nbsp;over weekend</p>
 
 
         </td>
 
         </td>
 
   </tr>
 
   </tr>
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   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Assembly <br/>Propagations</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Preparations <br/>Propagations</h3></div></td>
  
 
         <td class="white arrow" >
 
         <td class="white arrow" >
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   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
<td class="first rightline"><div class="heightdiv"><h3>Assembly <br/> Testing RBS's</h3></div></td>
+
<td class="first rightline"><div class="heightdiv"><h3>Testing RBS's <br/> Assembly</h3></div></td>
  
 
         <td class="white arrow" >
 
         <td class="white arrow" >
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   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
<td class="first rightline"><div class="heightdiv"><h3>Assembly <br/> Testing RBS's</h3></div></td>
+
<td class="first rightline"><div class="heightdiv"><h3></h3></div></td>
  
 
         <td class="white arrow" >
 
         <td class="white arrow" >
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         </td>
 
         </td>
 
   </tr>
 
   </tr>
  <tr>
 
        <td class="first rightline"><div class="heightdiv"><h3>Propagations</h3></div></td>
 
        <td class="white arrow" >
 
            <p style="font-size:13px;">Transform <br/>&nbsp;&nbsp;&nbsp; AH020 & AH021 <br/> into TOP10</p>
 
        </td>
 
        <td class="white arrow" >
 
            <p>Plates in rt <br/> over weekend</p>
 
        </td>
 
  </tr>
 
 
  
 
</table>
 
</table>
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   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Assembly <br/> Testing RBS's</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Testing RBS's <br/> Assembly</h3></div></td>
 
         <td class="white arrow" >
 
         <td class="white arrow" >
 
             <p>Re-assembling AH016 & <br/>AH018</p>
 
             <p>Re-assembling AH016 & <br/>AH018</p>
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   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Testing RBS's</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Testing RBS's <br/>Propagation</h3></div></td>
 
         <td class="white arrow" >
 
         <td class="white arrow" >
 
             <p style="font-size:12px">o/n cultures of <br/>&nbsp;AH016 & AH018 <br/> in TOP10</p>
 
             <p style="font-size:12px">o/n cultures of <br/>&nbsp;AH016 & AH018 <br/> in TOP10</p>
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   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Measurement <br/> Testing RBS's</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Testing RBS's <br/>Measurement</h3></div></td>
 
         <td class="white arrow" >
 
         <td class="white arrow" >
 
             <p>AH018 in <br/>BL21(DE3)</p>
 
             <p>AH018 in <br/>BL21(DE3)</p>
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   </tr>
 
   </tr>
 
     <tr>
 
     <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Producing linearized backbones</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Preparations <br/> Linearized Backbones</h3></div></td>
  
 
         <td class="white arrow" >
 
         <td class="white arrow" >
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   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Preparations<br/> Testing RBS's</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Preparations <br/> Linearized Backbones</h3></div></td>
 
         <td class="white arrow" >
 
         <td class="white arrow" >
 
             <p style="font-size:12px">Transform <br/>AH007-AH009 <br/> into TOP10</p>
 
             <p style="font-size:12px">Transform <br/>AH007-AH009 <br/> into TOP10</p>
Line 705: Line 694:
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Measurement <br/> Testing RBS's</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Testing RBS's <br/> Measurement</h3></div></td>
 
         <td class="white arrow" >
 
         <td class="white arrow" >
 
             <p style="font-size:12px">Refresh AH016 &<br/>AH018 on <br/> IPTG plate</p>
 
             <p style="font-size:12px">Refresh AH016 &<br/>AH018 on <br/> IPTG plate</p>
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   </tr>
 
   </tr>
 
     <tr>
 
     <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Violacein construct</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Violacein Plasmid <br/> Construction</h3></div></td>
 
        
 
        
 
         <td class="white arrow" >
 
         <td class="white arrow" >
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   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Constructing Cellulose system</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Preparations</h3></div></td>
 +
        <td class="white arrow" >
 +
            <p>Preparing <br/>&nbsp;&nbsp;PK BL21(DE3)<br/> competent cells</p>
 +
        </td>
 +
        <td class="blue arrow" >
 +
            <p>Cells stored <br/>in -80°C</p>
 +
        </td>       
 +
  </tr>
 +
  <tr>
 +
        <td class="first rightline"><div class="heightdiv"><h3>Cellulose Plasmid <br/> Construction</h3></div></td>
 
         <td class="white arrow" >
 
         <td class="white arrow" >
 
             <p>PCR AH021 <br/>with <br/>P019&P020</p>
 
             <p>PCR AH021 <br/>with <br/>P019&P020</p>
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             <p>Transformants <br/>stored in <br/>+4°C</p>
 
             <p>Transformants <br/>stored in <br/>+4°C</p>
 
         </td>
 
         </td>
  </tr>
 
  <tr>
 
        <td class="first rightline"><div class="heightdiv"><h3>Preparations</h3></div></td>
 
        <td class="white arrow" >
 
            <p>Preparing <br/>&nbsp;&nbsp;PK BL21(DE3)<br/> competent cells</p>
 
        </td>
 
        <td class="blue arrow" >
 
            <p>Cells stored <br/>in -80°C</p>
 
        </td>       
 
 
   </tr>
 
   </tr>
 
     <tr>
 
     <tr>
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</tr>
 
</tr>
 
     <tr>
 
     <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Violacein Construct</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Violacein Plasmid <br/> Construction</h3></div></td>
 
              
 
              
 
         </td>
 
         </td>
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   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Preparations</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Preparations <br/> <span style="font-size:20px">Propagations</span></h3></div></td>
  
 
         <td class="white arrow" >
 
         <td class="white arrow" >
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   <tr>
 
   <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Propane Plasmids 1 & 2</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Propane Plasmids 1&2 <span style="font-size:20px">Construction </span></h3></div></td>
 
          
 
          
 
<td class="white arrow" >
 
<td class="white arrow" >
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   <tr>
 
   <tr>
         <td class="first rightline"><div class="heightdiv"><h3>GFP brick</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>GFP Brick <span style="fonst-size:20px">Construction</span></h3></div></td>
  
 
<td class="white arrow" >
 
<td class="white arrow" >
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   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Propane Plasmid 2</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Propane Plasmid 2 <span style="font-size:20px">Construction</span></h3></div></td>
  
 
<td class="white arrow" >
 
<td class="white arrow" >
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<h2 id="week10">Week 10</h2>
 
<h2 id="week10">Week 10</h2>
 +
 +
 
<h2 id="week11">Week 11</h2>
 
<h2 id="week11">Week 11</h2>
  
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   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Constructing Propane Plasmid 1</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Propane Plasmid 1 <span style="font-size:20px">Construction</span></h3></div></td>
 
         <td class="red arrow" >
 
         <td class="red arrow" >
 
             <p>Continue CAR1 <br/>colony <br/>screening</p>
 
             <p>Continue CAR1 <br/>colony <br/>screening</p>
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   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Preparations</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Preparations <span style="font-size:20px">Linearized Backbones</span></h3></div></td>
 
         <td class="white arrow" >
 
         <td class="white arrow" >
 
             <p>Re-restrict <br/>AH043</p>
 
             <p>Re-restrict <br/>AH043</p>
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   </tr>
 
   </tr>
 
     <tr>
 
     <tr>
         <td class="first rightline"><div class="heightdiv"><h3></h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3> <span style="font-size:20px">Propagations</span></h3></div></td>
 
              
 
              
 
         <td class="white arrow" >
 
         <td class="white arrow" >
Line 1,023: Line 1,014:
 
</tr>
 
</tr>
 
     <tr>
 
     <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Constructing Propane Plasmid 2</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Propane Plasmid 2 <span style="font-size:20px">Construction</span></h3></div></td>
 
              
 
              
 
         </td>
 
         </td>
Line 1,037: Line 1,028:
 
</tr>
 
</tr>
 
     <tr>
 
     <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Constructing Cellulose Plasmid</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Cellulose Plasmid <span style="font-size:20px">Construction</span></h3></div></td>
 
              
 
              
 
         <td class="red arrow" >
 
         <td class="red arrow" >
Line 1,060: Line 1,051:
 
</tr>
 
</tr>
 
     <tr>
 
     <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Constructing Propane Plasmid 1</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3></h3></div></td>
 +
       
 +
        <td class="white arrow" >
 +
            <p>2nd PCR <br/> for BglX <br/> (from AH021)</p>
 +
        </td>
 +
<td class="white arrow" >
 +
            <p>Stored in -4°C <br/>over the <br/>weekend</p>
 +
        </td>
 +
</tr>
 +
  <tr>
 +
        <td class="first rightline"><div class="heightdiv"><h3>Propane Plasmid 1 <span style="font-size:20px">Construction</span></h3></div></td>
 
              
 
              
 
         </td>
 
         </td>
Line 1,084: Line 1,085:
 
         </td>
 
         </td>
 
</tr>
 
</tr>
 +
 
     <tr>
 
     <tr>
         <td class="first rightline"><div class="heightdiv"><h3>Constructing Cellulose Plasmid</h3></div></td>
+
         <td class="first rightline"><div class="heightdiv"><h3>Violacein Plasmid <span style="font-size:2px">Construction</span></h3></div></td>
       
+
        <td class="white arrow" >
+
            <p>2nd PCR <br/> for BglX <br/> (from AH021)</p>
+
        </td>
+
<td class="white arrow" >
+
            <p>Stored in -4°C <br/>over the <br/>weekend</p>
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        <td class="first rightline"><div class="heightdiv"><h3>Constructing Violacein construct</h3></div></td>
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Revision as of 13:57, 12 August 2015

Lab Book

An example of what the table could look like:

Construct
worked with

Things done

Lab 1.1.2015
Monday

CAR

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text, there
is lot of it

another CAR?

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Here is my
text, there
is lot of it

Ado?

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Ado cont.

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Here is my
text, there
is lot of it

Week 1

Construct
worked with

Things done

May 18th -
May 22nd

Preparations
Plates

Making Cm
plates

Stored in
+4°C

Preparations
Competent Cells

Preparing
solutions for
competent cells

Stored in
+4°C

o/n culture
of E. coli
TOP10

Chemical
competent
cells protocol

Tranformation
efficiency kit

o/n culture
of E. coli
BL21(DE3)

Chemical
competent
cells protocol

Tranformation
efficiency kit

Colonies are
the wrong color

Preparations
Propagations

Transformed
  AH001-AH008
into TOP10

 AH001-AH004
& AH006

Stored in
+4°C

No colonies for
   AH005, AH007
or AH008

Week 2

Construct
worked with

Things done

May 25th -
May 29th

Preparations
Propagations

Transformed
AH007-AH009,
AH005 & AH019
into TOP10

AH007-AH009
& AH019

o/n cultures

MiniPrep

o/n culture of
 AH001-AH004
& AH006

MiniPrep

No colonies
for AH005

Re-transform
into TOP10

Plates in rt
over weekend

Checking
Competent Cells

o/n culture
of BL21(DE3)
transformants

MiniPrep

PCR with
VF2 & VR

  Smeared gels

AH001+
   AH003+AH008
-> AH013

Repeat
ligation

Restrict &
extract
from gel

 Transformation
into TOP10

Plates in rt
over weekend

Testing RBS's
Assembly

Restrict
AH001-AH004
& AH006-AH008

Ligation

AH002+
  AH004+AH008
-> AH014

Repeat
ligation

Restrict &
extract
from gel

 Transformation
into TOP10

Plates in rt
over weekend

AH001+
   AH006+AH008
-> AH011

Repeat
ligation

Restrict &
extract
from gel

 Transformation
into TOP10

Plates in rt
over weekend

Week 3

Construct
worked with

Things done

June 1st -
June 5th

Preparations
Propagations

Transform
    AH020 & AH021
into TOP10

Plates in rt
 over weekend

Testing RBS's
Assembly

No colonies
for AH011
or AH014

re-assemble

Restrict &
extract
from gel

Ligate &
tranform
into TOP10

No colonies

AH005 &
AH013 OK

o/n cultures

MiniPrep

Week 4

Construct
worked with

Things done

June 8th -
June 12th

Preparations
Propagations

o/n cultures
for AH020
& AH021

MiniPrep

Testing RBS's
Assembly

Re-assembling
  AH011, AH013
& AH014

Transform into TOP10

o/n cultures

MinPrep

Assembling
AH015

  Ligate AH002+
  AH005+AH008

Restrict &
extract
from gel

Transform into TOP10

o/n cultures

MiniPrep

Week 5

Week 6

Construct
worked with

Things done

June 22nd -
June 26th

Testing RBS's
Assembly

Re-assembling AH016 &
AH018

Transform
into
BL21(DE3)

No colonies

Testing RBS's
Propagation

o/n cultures of
 AH016 & AH018
in TOP10

MiniPrep

Transform
into
BL21(DE3)

Plates in rt
over the
weekend

Testing RBS's
Measurement

AH018 in
BL21(DE3)

o/n culture

IPTG induction &
    blue chromoprotein
measurement

No blue chromoprotein

Refresh on
IPTG plate

No blue chromoprotein

Preparations
Linearized Backbones

o/n cultures of
AH007-AH009

MiniPrep

Restrict &
extract
from gel

Week 7

Construct
worked with

Things done

June 29th -
July 3rd

Preparations
Linearized Backbones

Transform
AH007-AH009
into TOP10

No colonies
(mistake in
protocol)

Use iGEM's
linearized
backbones

Testing RBS's

o/n cultures of
  AH016&AH018
in TOP10

IPTG
induction

No Blue
chromoprotein

Testing RBS's
Measurement

Refresh AH016 &
AH018 on
IPTG plate

No blue
chromoprotein

Violacein Plasmid
Construction

PCR AH20
with
P012&P013

PCR
purification

VioA waits
for 2nd PCR

PCR AH20
with
P024&P025

PCR
unsuccesful

PCR AH20
with
P034&P035

PCR
purification

VioE waits
for 2nd PCR

Week 8

Construct
worked with

Things done

July 6th -
July 10th

Preparations

Preparing
  PK BL21(DE3)
competent cells

Cells stored
in -80°C

Cellulose Plasmid
Construction

PCR AH021
with
P019&P020

Problem with
PCR machine

Re-do PCR

PCR
purifcation

BglX waits for
2nd PCR

AH029 into
TOP10

Transformants
stored in
+4°C

Testing RBS's

Transform AH016
& AH018 into
PK BL21(DE3)

Refresh
transformants
on IPTG plates

No blue
chromoprotein

Violacein Plasmid
Construction

Gradient PCR for
AH020 with
P024&P025

PCR
purification

Week 9

Construct
worked with

Things done

July 13th -
July 17th

Preparations
Propagations

o/n culture
of AH029

MiniPrep

CAR2 OK

PCR
purification

Propane Plasmids 1&2 Construction

PCR for
CAR1-3 &
AtoB1-2

CAR1,3 &
AtoB1-2
unspecific bands

Gradient PCR
for CAR3 & 4

PCR with DMSO
for CAR1, 3
& AtoB1, 2

dunnowhat

GFP Brick Construction

o/n culture
of AH044

MiniPrep

Propane Plasmid 2 Construction

PCR for
AtoB3-4

Unspecific
bands

AtoB 4
extracted
from gel

Low
concentration

Gradient
PCR for
AtoB3

Extract &
purify
from gel

Week 10

Week 11

Construct
worked with

Things done

July 27th -
July 31st

Propane Plasmid 1 Construction

Continue CAR1
colony
screening

Screen more
colonies

PCR purification

PCR for
new CAR2

Gradient PCR
with KAPA
polymerase

PCR
purification

Preparations Linearized Backbones

Re-restrict
AH043

Gel purification

Low yield

Re-restrict

Propagations

o/n culture
of AH031

MiniPrep

Transform
AH033 into
TOP10

o/n culture

MiniPrep

Low yield

o/n culture

MiniPrep

Low yield,
but stored
in -20°C

Propane Plasmid 2 Construction

TOPO Blunt End
Cloning for AtoB1
and transformation
into TOP10

Screening
with colony
PCR

PCR
purification

Cellulose Plasmid Construction

PCR for
Cen1 & Cen2

Gradient PCR
with KAPA
polymerase

PCR
purification for
Cen2

Gel purification
for Cen1

2nd PCR
for BglX
(from AH021)

Stored in -4°C
over the
weekend

Propane Plasmid 1 Construction

Re-do colony
PCR for CAR3
successful TOPO colony

Re-do
colony PCR

o/n culture of
the colony

MiniPrep

PCR

Violacein Plasmid Construction

2nd PCR
for VioA
(from AH020)

Stored in -4°C
over the
weekend

KAPA gradient
PCR for AtoB3,4
& CAR3

Gel purification
for AtoB4 & Car3

PCR
purification
for AtoB3

Week 12

Week 13

Week 14

Week 15

Week 16

Week 17

Week 18