Team:Northwestern/CFPS

From 2014.igem.org

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<h5>20140723 Testing our lysates</h5>
<h5>20140723 Testing our lysates</h5>
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<li>data from BL21 (E.Coli) lysates</li>
<li>data from BL21 (E.Coli) lysates</li>
<li>out of K12, DH5a, and BL21, BL21 expressed the most fluorescence</li>
<li>out of K12, DH5a, and BL21, BL21 expressed the most fluorescence</li>
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<li>the plasmid used was superfolder GFP from Jewett Lab</li>
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<li>the plasmid used was a construct of superfolder GFP from the Jewett Lab</li>
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<li><b>conclusion:</b>our lysates are functional</li>
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<img src="https://static.igem.org/mediawiki/2014/0/05/072314CFPS.png" alt="red: control, blue: BL21 lysate"/>
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<img class="img-rounded" src="https://static.igem.org/mediawiki/2014/6/65/E.coli_lysates.jpg" style="width:100px; padding-bottom: 20px;"/>
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<img class="img-rounded" src="https://static.igem.org/mediawiki/2014/3/32/E.coli_lysates_2.jpg" style="width:100px;"/>
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<h5>20140801 Making our own mastermix (iMM)!</h5>
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<ul>
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<li>RM: Rey's mastermix</li>
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<li>iMM: we made our own according to Rey's recipe</li>
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<li>looks like our mastermix works fine</li>
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<li><b>conclusion</b>: we will use our own mastermix from now on</li>
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<li>the second part: we made s. rimosus lysates</li>
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<li>we used a sfgfp construct from the Jewett lab</li>
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<li>the lysates seem to express some gfp!</li>
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<iframe width="768" height="346" seamless frameborder="0" scrolling="no" src="https://docs.google.com/spreadsheets/d/1UTZ4xUKjVp4DbYd82_U266bu_I0fe3BVjfnmZkRn5t4/pubchart?oid=1393566635&amp;format=interactive"></iframe>
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<img src="https://static.igem.org/mediawiki/2014/5/52/IMMRMBL21.jpg"style="width: 800px; padding-bottom:40px;"/>
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<img src="https://static.igem.org/mediawiki/2014/1/1b/Srsfgfp.jpg"style="width: 800px; padding-bottom: 40px;"/>
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<h5>20140915 All of our lysates</h5>
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<ul>
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<li>RBS B0034</li>
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<li>Tried all of our lysates</li>
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<li>green fluorescence indicates mRNA levels of transcription</li>
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<li>red fluorescence indicates protein levels of translation</li>
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<li><b>conclusion</b>: the data is most likely statistically insignificant</li>
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</ul>
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<img src="https://static.igem.org/mediawiki/2014/9/94/RFP091514.png"style="width: 800px;"/>
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<img src="https://static.igem.org/mediawiki/2014/c/cf/GFP_091514.png"style="width: 800px;"/>
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<h5>20141014 Cell-Free Protein Synthesis Assay with All of our Lysates</h5>
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<ul>
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<li>We used RBS No. 5 (B0034) with RFP-Spinach Aptamer</li>
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<li>Displayed here is the first read</li>
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<li>The kinetic portion of the plate reader did not record data in all of our wells</li>
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<li>We will continue doing CFPS experiments </li>
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<li><b>conclusion</b>: we need more time! Looks like fluorophores were incorrectly added</li>
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</ul>
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<img src="https://static.igem.org/mediawiki/2014/f/f2/Otherlysatesoctober.jpg"style="width: 800px; padding-bottom: 40px;"/>
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<img src="https://static.igem.org/mediawiki/2014/4/4a/E.colioctober.jpg"style="width: 800px; padding-bottom: 40px;"/>
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<img src="https://static.igem.org/mediawiki/2014/0/00/Spinapt101514.png" style="width: 800px; padding-bottom: 40px;"/>
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<img src="https://static.igem.org/mediawiki/2014/c/c0/Jewettlabsfgfp.png" style="width: 800px; padding-bottom: 40px;"/>
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<img src="https://static.igem.org/mediawiki/2014/c/cf/RFPE.Coli101514.png" style="width: 800px; padding-bottom: 40px;"/>
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Latest revision as of 23:19, 17 October 2014

Dropdown menu from bootstrap

Cell-Free Protein Synthesis

20140723 Testing our lysates
  • data from BL21 (E.Coli) lysates
  • out of K12, DH5a, and BL21, BL21 expressed the most fluorescence
  • the plasmid used was a construct of superfolder GFP from the Jewett Lab
  • conclusion:our lysates are functional
red: control, blue: BL21 lysate
20140801 Making our own mastermix (iMM)!
  • RM: Rey's mastermix
  • iMM: we made our own according to Rey's recipe
  • looks like our mastermix works fine
  • conclusion: we will use our own mastermix from now on
  • the second part: we made s. rimosus lysates
  • we used a sfgfp construct from the Jewett lab
  • the lysates seem to express some gfp!
20140915 All of our lysates
  • RBS B0034
  • Tried all of our lysates
  • green fluorescence indicates mRNA levels of transcription
  • red fluorescence indicates protein levels of translation
  • conclusion: the data is most likely statistically insignificant
20141014 Cell-Free Protein Synthesis Assay with All of our Lysates
  • We used RBS No. 5 (B0034) with RFP-Spinach Aptamer
  • Displayed here is the first read
  • The kinetic portion of the plate reader did not record data in all of our wells
  • We will continue doing CFPS experiments
  • conclusion: we need more time! Looks like fluorophores were incorrectly added

Retrieved from "http://2014.igem.org/Team:Northwestern/CFPS"