Difference between revisions of "Team:Pittsburgh/Notebook"

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<li><a href="#Week5" class="table">Week 5: June 20 - June 26</a></li>
 
<li><a href="#Week5" class="table">Week 5: June 20 - June 26</a></li>
 
<li><a href="#Week6" class="table">Week 6: June 27 - July 3</a></li>
 
<li><a href="#Week6" class="table">Week 6: June 27 - July 3</a></li>
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<li><a href="#Week7" class="table">Week 7: July 5 - July 8</a></li>
 
</ul>
 
</ul>
 
</div>
 
</div>
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<a href="https://static.igem.org/mediawiki/2016/7/7c/TeamPittsburghNotebookWeek5.pdf" target="_blank">Week 5 Notebook</a><br>
 
<a href="https://static.igem.org/mediawiki/2016/7/7c/TeamPittsburghNotebookWeek5.pdf" target="_blank">Week 5 Notebook</a><br>
 
<a href="#Top">Back to Top</a>
 
<a href="#Top">Back to Top</a>
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<h1 class="nav"><a name="Week6" class="nav">Week 6: June 27 - July 3</a></h1>
 
<h1 class="nav"><a name="Week6" class="nav">Week 6: June 27 - July 3</a></h1>
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</ul>
 
</ul>
 
<a href="https://static.igem.org/mediawiki/2016/5/59/T--Pittsburgh--NotebookWeek6.pdf" target="_blank">Week 6 Notebook</a><br>
 
<a href="https://static.igem.org/mediawiki/2016/5/59/T--Pittsburgh--NotebookWeek6.pdf" target="_blank">Week 6 Notebook</a><br>
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<a href="#Top">Back to Top</a>
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<h1 class="nav"><a name="Week7" class="nav">Week 7: July 5 - July 8</a></h1>
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<h2>Wet Lab</h2>
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<h3>Reporter</h3>
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<ul>
 +
    <li>Sequenced amilCP construct does not contain amilCP</li>
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    <li>Unsuccessfully linearize and amplify eGFP construct using <a href="https://2016.igem.org/Team:Pittsburgh/Protocols#pcr" target="_blank">PCR</a></li>
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</ul>
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<h3>Cell-Free Extract</h3>
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    <ul>
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        <li>384-well plate requires at least 10 μL of reaction</li>
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    </ul>
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<h3>DNAzyme</h3>
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    <ul>
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        <li><a href="https://2016.igem.org/Team:Pittsburgh/Protocols#annealing" target="_blank">Anneal</a> PO strand with catalytic strand, both with and without erbium</li>
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        <li>Test success of annealing reaction in cell-free extract and with acrylamide gels</li>
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    </ul>
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<h2>Dry Lab</h2>
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<ul>
 +
<li>Practice outreach presentation for tissue engineering camp</li>
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<li>Develop DNAzymes for other heavy metals</li>
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</ul>
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<a href="https://static.igem.org/mediawiki/2016/d/d7/T--Pittsburgh--NotebookWeek7.pdf" target="_blank">Week 7 Notebook</a><br>
 
<a href="#Top">Back to Top</a>
 
<a href="#Top">Back to Top</a>
 
      
 
      

Revision as of 17:01, 10 July 2016

Thank you to our sponsors!

Office of the Provost Department of Bioengineering Department of Biological Sciences Department of Chemistry Swanson School of Engineering
IDT NEB Experiment

Our weekly progress. For a list of our protocols, visit the Protocols page

Contents

Week 1: May 23 - May 27

Wet Lab

Dry Lab

  • Brainstorm genetic circuits for a thallium sensor
  • Lab safety training
Week 1 Notebook
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Week 2: May 31 - June 3

Wet Lab

Cell-Free Extract

Dry Lab

  • Contact museums and summer programs for outreach opportunities
  • Lab safety training
Week 2 Notebook
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Week 3: June 6 - June 12

Wet Lab

Reporter

  • Transform T7 promoter, amilCP, and terminator
  • Begin assembly by ligating linearized T7 promoter and amilCP

Dry Lab

  • Contact museums and summer programs for outreach opportunities
Week 3 Notebook
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Week 4: June 13 - June 17

Wet Lab

Reporter

  • Transform T7-GFP plasmid, lacZ alpha fragment, and eGFP
  • Send promising T7 promoter -- amilCP ligations to be sequenced
  • Perform double digest of T7 promoter and terminator from last week
  • Ligate double-digested T7 promoter to new reporters (lacZ and eGFP)

Dry Lab

  • TECBio, DiSCoBio, and Tissue Engineering Camp outreach opportunities set
Week 4 Notebook
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Week 5: June 20 - June 26

Wet Lab

Reporter

  • Ligate T7 promoter -- amilCP construct to terminator
  • Extract successful ligations of T7 promoter to eGFP
  • Ligate T7 promoter -- eGFP construct to terminator

Cell-Free Extract

  • Reaction volume can be reduced to 5 μL with 5 ng/μL of DNA

Toehold Switch

  • Collins triggers activate the switches (both in plasmid form) to express LacZ

Dry Lab

  • Reach out to teams to collaborate based on last year's projects
Week 5 Notebook
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Week 6: June 27 - July 3

Wet Lab

Reporter

  • Identify successful ligations to terminator for amilCP and eGFP consturcts using a gel
  • Send correct plasmids for sequencing for confirmation
  • Test plasmids in cell-free extract
  • amilCP does not produce color in cell-free reaction
  • eGFP produces fluorescence comparable to that from the Collins T7-GFP plasmid in cell-free reaction
  • Linearized plasmids containing only the promoter and insert (no terminator) do not express protein

Cell-Free Extract

  • Reaction volume can be reduced to 1 μL with 5 ng/μL of DNA

Toehold Switch

  • Collins plasmids express LacZ with 25 ng of switch
  • DNA oligos trigger Collins switches

Dry Lab

  • Work on outreach presentation for tissue engineering camp
Week 6 Notebook
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Week 7: July 5 - July 8

Wet Lab

Reporter

  • Sequenced amilCP construct does not contain amilCP
  • Unsuccessfully linearize and amplify eGFP construct using PCR

Cell-Free Extract

  • 384-well plate requires at least 10 μL of reaction

DNAzyme

  • Anneal PO strand with catalytic strand, both with and without erbium
  • Test success of annealing reaction in cell-free extract and with acrylamide gels

Dry Lab

  • Practice outreach presentation for tissue engineering camp
  • Develop DNAzymes for other heavy metals
Week 7 Notebook
Back to Top