Difference between revisions of "Team:Calgary/Protocols"

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                                 </td>
 
                                 </td>
 
                                 <td>
 
                                 <td>
−
                                     <p>Registry DNA was rehydrated for completion of the Interlab Study. Also,
+
                                     <p>This section might be deleted.</p>
−
                                        Part:BBa_K934001 (phaC1-A-B1) was rehydrated and transformed into our chassis
+
−
                                        so that PHB was produced and preliminary secretion assays could be performed
+
−
                                        before the Synthesis subgroup had completed their cloning.</p>
+
 
                                 </td>
 
                                 </td>
 
                             </tr>
 
                             </tr>
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                                 <td>
 
                                 <td>
 
                                     <ul>
 
                                     <ul>
−
                                         <li>iGEM 2017 distribution kit</li>
+
                                         <li>iGEM 2018 distribution kit</li>
 
                                         <li>ddH₂O</li>
 
                                         <li>ddH₂O</li>
 
                                     </ul>
 
                                     </ul>
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                             <tr>
 
                             <tr>
 
                                 <td>
 
                                 <td>
−
                                     <h5>Protocols</h5>
+
                                     <h5>Protocol</h5>
 
                                 </td>
 
                                 </td>
 
                                 <td>
 
                                 <td>
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                                         <li>Pipette up and down 3-5 times.</li>
 
                                         <li>Pipette up and down 3-5 times.</li>
 
                                         <li>Incubate at room temperature for 10 minutes.</li>
 
                                         <li>Incubate at room temperature for 10 minutes.</li>
−
                                         <li>Transform cells with 1μL of rehydrated DNA. Store the remaining amount at
+
                                         <li>Transform cells with 1μL of rehydrated DNA as per transformation protocol. Store the remaining amount at
 
                                             -20°C.</li>
 
                                             -20°C.</li>
 
                                     </ol>
 
                                     </ol>
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                         <i id="icon2" class="fas fa-plus accordion-icon" aria-hidden="true"></i>
 
                         <i id="icon2" class="fas fa-plus accordion-icon" aria-hidden="true"></i>
 
                         <h3>
 
                         <h3>
−
                             Protocol 2
+
                             Rehydration of Synthesized DNA
 
                         </h3>
 
                         </h3>
 
                     </div>
 
                     </div>
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                                 </td>
 
                                 </td>
 
                                 <td>
 
                                 <td>
−
                                     <p>Registry DNA was rehydrated for completion of the Interlab Study. Also,
+
                                     <p>This section might be deleted.</p>
−
                                        Part:BBa_K934001 (phaC1-A-B1) was rehydrated and transformed into our chassis
+
−
                                        so that PHB was produced and preliminary secretion assays could be performed
+
−
                                        before the Synthesis subgroup had completed their cloning.</p>
+
 
                                 </td>
 
                                 </td>
 
                             </tr>
 
                             </tr>
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                                 <td>
 
                                 <td>
 
                                     <ul>
 
                                     <ul>
−
                                         <li>iGEM 2017 distribution kit</li>
+
                                         <li>Synthesized DNA from IDT or Genscript</li>
 
                                         <li>ddH₂O</li>
 
                                         <li>ddH₂O</li>
 
                                     </ul>
 
                                     </ul>
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                             <tr>
 
                             <tr>
 
                                 <td>
 
                                 <td>
−
                                     <h5>Protocols</h5>
+
                                     <h5>Protocol *NEED TO VERIFY*</h5>
 
                                 </td>
 
                                 </td>
 
                                 <td>
 
                                 <td>
 
                                     <ol>
 
                                     <ol>
−
                                         <li>Add 10μL of ddH₂O to the desired well.</li>
+
                                         <li>Centrifuge tube containing the synthesized DNA for 3-5 seconds at 3000g to ensure that all material is at the bottom of the tube.</li>
−
                                         <li>Pipette up and down 3-5 times.</li>
+
                                        <li>Add ddH₂O to reach a final concentration of 50 ng/μL.</li>
−
                                         <li>Incubate at room temperature for 10 minutes.</li>
+
                                         <li>Vortex.</li>
−
                                        <li>Transform cells with 1μL of rehydrated DNA. Store the remaining amount at
+
                                         <li>Incubate at 50°C for 20 minutes.</li>
−
                                            -20°C.</li>
+
                                      <li>Briefly vortex and centrifuge. Store at -20°C.</li>
 
                                     </ol>
 
                                     </ol>
 
                                 </td>
 
                                 </td>

Revision as of 22:31, 15 October 2018

Team:Calgary/Notebook - 2018.igem.org

PROTOCOLS



Below are the protocols used by the team.

Experimental Details

This section might be deleted.
Materials
  • iGEM 2018 distribution kit
  • ddH₂O
Protocol
  1. Add 10μL of ddH₂O to the desired well.
  2. Pipette up and down 3-5 times.
  3. Incubate at room temperature for 10 minutes.
  4. Transform cells with 1μL of rehydrated DNA as per transformation protocol. Store the remaining amount at -20°C.

Experimental Details

This section might be deleted.
Materials
  • Synthesized DNA from IDT or Genscript
  • ddH₂O
Protocol *NEED TO VERIFY*
  1. Centrifuge tube containing the synthesized DNA for 3-5 seconds at 3000g to ensure that all material is at the bottom of the tube.
  2. Add ddH₂O to reach a final concentration of 50 ng/μL.
  3. Vortex.
  4. Incubate at 50°C for 20 minutes.
  5. Briefly vortex and centrifuge. Store at -20°C.

Experimental Details

Registry DNA was rehydrated for completion of the Interlab Study. Also, Part:BBa_K934001 (phaC1-A-B1) was rehydrated and transformed into our chassis so that PHB was produced and preliminary secretion assays could be performed before the Synthesis subgroup had completed their cloning.
Materials
  • iGEM 2017 distribution kit
  • ddH₂O
Protocols
  1. Add 10μL of ddH₂O to the desired well.
  2. Pipette up and down 3-5 times.
  3. Incubate at room temperature for 10 minutes.
  4. Transform cells with 1μL of rehydrated DNA. Store the remaining amount at -20°C.

Experimental Details

Registry DNA was rehydrated for completion of the Interlab Study. Also, Part:BBa_K934001 (phaC1-A-B1) was rehydrated and transformed into our chassis so that PHB was produced and preliminary secretion assays could be performed before the Synthesis subgroup had completed their cloning.
Materials
  • iGEM 2017 distribution kit
  • ddH₂O
Protocols
  1. Add 10μL of ddH₂O to the desired well.
  2. Pipette up and down 3-5 times.
  3. Incubate at room temperature for 10 minutes.
  4. Transform cells with 1μL of rehydrated DNA. Store the remaining amount at -20°C.

Experimental Details

Registry DNA was rehydrated for completion of the Interlab Study. Also, Part:BBa_K934001 (phaC1-A-B1) was rehydrated and transformed into our chassis so that PHB was produced and preliminary secretion assays could be performed before the Synthesis subgroup had completed their cloning.
Materials
  • iGEM 2017 distribution kit
  • ddH₂O
Protocols
  1. Add 10μL of ddH₂O to the desired well.
  2. Pipette up and down 3-5 times.
  3. Incubate at room temperature for 10 minutes.
  4. Transform cells with 1μL of rehydrated DNA. Store the remaining amount at -20°C.

Experimental Details

Registry DNA was rehydrated for completion of the Interlab Study. Also, Part:BBa_K934001 (phaC1-A-B1) was rehydrated and transformed into our chassis so that PHB was produced and preliminary secretion assays could be performed before the Synthesis subgroup had completed their cloning.
Materials
  • iGEM 2017 distribution kit
  • ddH₂O
Protocols
  1. Add 10μL of ddH₂O to the desired well.
  2. Pipette up and down 3-5 times.
  3. Incubate at room temperature for 10 minutes.
  4. Transform cells with 1μL of rehydrated DNA. Store the remaining amount at -20°C.