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Team:Northwestern/CFPS
From 2014.igem.org
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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> | ||
| - | <li>the plasmid used was superfolder GFP from Jewett Lab</li> | + | <li>the plasmid used was a construct of superfolder GFP from the Jewett Lab</li> |
| - | <li><b>conclusion</b>our lysates are | + | <li><b>conclusion:</b>our lysates are functional</li> |
</ul> | </ul> | ||
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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;"/> | <img class="img-rounded" src="https://static.igem.org/mediawiki/2014/3/32/E.coli_lysates_2.jpg" style="width:100px;"/> | ||
</div> | </div> | ||
| + | </div><!--row--> | ||
| + | |||
| + | <div class="row"> | ||
| + | <div class="col-md-3"> | ||
| + | <h5>20140801 Making our own mastermix (iMM)!</h5> | ||
| + | <ul> | ||
| + | <li>RM: Rey's mastermix</li> | ||
| + | <li>iMM: we made our own according to Rey's recipe</li> | ||
| + | <li>looks like our mastermix works fine</li> | ||
| + | <li><b>conclusion</b>: we will use our own mastermix from now on</li> | ||
| + | <li>the second part: we made s. rimosus lysates</li> | ||
| + | <li>we used a sfgfp construct from the Jewett lab</li> | ||
| + | <li>the lysates seem to express some gfp!</li> | ||
| + | </ul> | ||
| + | |||
| + | </div> | ||
| + | <div class="col-md-9"> | ||
| + | <img src="https://static.igem.org/mediawiki/2014/5/52/IMMRMBL21.jpg"style="width: 800px; padding-bottom:40px;"/> | ||
| + | |||
| + | <img src="https://static.igem.org/mediawiki/2014/1/1b/Srsfgfp.jpg"style="width: 800px; padding-bottom: 40px;"/> | ||
| + | </div> | ||
| + | |||
</div><!--row--> | </div><!--row--> | ||
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<li>RBS B0034</li> | <li>RBS B0034</li> | ||
<li>Tried all of our lysates</li> | <li>Tried all of our lysates</li> | ||
| - | <li>green fluorescence indicates | + | <li>green fluorescence indicates mRNA levels of transcription</li> |
| - | <li>red fluorescence indicates | + | <li>red fluorescence indicates protein levels of translation</li> |
| - | <li><b>conclusion</b>: the data is most likely statistically | + | <li><b>conclusion</b>: the data is most likely statistically insignificant</li> |
</ul> | </ul> | ||
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</div><!--row--> | </div><!--row--> | ||
| + | |||
| + | |||
<div class="row"> | <div class="row"> | ||
<div class="col-md-3"> | <div class="col-md-3"> | ||
| - | <h5>20141014 | + | <h5>20141014 Cell-Free Protein Synthesis Assay with All of our Lysates</h5> |
<ul> | <ul> | ||
| - | <li>We used RBS | + | <li>We used RBS No. 5 (B0034) with RFP-Spinach Aptamer</li> |
<li>Displayed here is the first read</li> | <li>Displayed here is the first read</li> | ||
<li>The kinetic portion of the plate reader did not record data in all of our wells</li> | <li>The kinetic portion of the plate reader did not record data in all of our wells</li> | ||
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</div> | </div> | ||
<div class="col-md-9"> | <div class="col-md-9"> | ||
| - | <img src="https://static.igem.org/mediawiki/2014/f/f2/Otherlysatesoctober.jpg"style="width: 800px; | + | <img src="https://static.igem.org/mediawiki/2014/f/f2/Otherlysatesoctober.jpg"style="width: 800px; padding-bottom: 40px;"/> |
| - | <img src="https://static.igem.org/mediawiki/2014/4/4a/E.colioctober.jpg"style="width: 800px;"/> | + | <img src="https://static.igem.org/mediawiki/2014/4/4a/E.colioctober.jpg"style="width: 800px; padding-bottom: 40px;"/> |
| + | <img src="https://static.igem.org/mediawiki/2014/0/00/Spinapt101514.png" style="width: 800px; padding-bottom: 40px;"/> | ||
| + | <img src="https://static.igem.org/mediawiki/2014/c/c0/Jewettlabsfgfp.png" style="width: 800px; padding-bottom: 40px;"/> | ||
| + | <img src="https://static.igem.org/mediawiki/2014/c/cf/RFPE.Coli101514.png" style="width: 800px; padding-bottom: 40px;"/> | ||
</div> | </div> | ||
Latest revision as of 23:19, 17 October 2014
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
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
