Difference between revisions of "Team:Pittsburgh/Experiments"

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<ul class="table">
 
<ul class="table">
 
     <li><a href="#reporter" class="table">Reporter</a></li>
 
     <li><a href="#reporter" class="table">Reporter</a></li>
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    <li><a href="#cell-free" class="table">Cell-Free System</a></li>
 
     <li><a href="#DNAzyme" class="table">DNAzyme</a></li>
 
     <li><a href="#DNAzyme" class="table">DNAzyme</a></li>
 
     <li><a href="#toehold" class="table">Toehold Switch</a></li>
 
     <li><a href="#toehold" class="table">Toehold Switch</a></li>
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<div class="notebook column full_size">
 
<div class="notebook column full_size">
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    <p>For our daily activities and experiments, please visit our <a href="2016.igem.org/Team:Pittsburgh/Notebook" target="_blank">Notebook</a>. For a list of our protocols, please visit our <a href="2016.igem.org/Team:Pittsburgh/Protocols" target="_blank">Protocols</a> page. </p>
 
<h2><a name="reporter" class="nav">Reporter</a></h2>
 
<h2><a name="reporter" class="nav">Reporter</a></h2>
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    <h3>amilCP</h3>
 +
    <p>amilCP is a chromoprotein added to the iGEM Registry by Uppsala Sweden, 2012. We were unable to produce amilCP in bacteria or in our cell-free system.</p>
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    <a href="#top">Back to Top</a>
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<h2><a name="cell-free" class="nav">Cell-Free System</a></h2>
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    <h3>Linear versus Plasmid DNA</h3>
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    <p>We have good data from adding plasmids to our cell-free system. However, because the <a href="http://www.cell.com/abstract/S0092-8674(14)01291-4" target="_blank">Collins paper</a> specified the use of linear DNA in their cell-free systems, we compared the signal from GFP from linear and plasmid DNA. There is no siginificant difference between the two constructs.</p>
 +
    <h3>dilution</h3>
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    <h3>volume optimization</h3>
 
     <a href="#top">Back to Top</a>
 
     <a href="#top">Back to Top</a>
 
<h2><a name="DNAzyme" class="nav">DNAzyme</a></h2>
 
<h2><a name="DNAzyme" class="nav">DNAzyme</a></h2>
 
     <a href="#top">Back to Top</a>
 
     <a href="#top">Back to Top</a>
 
<h2><a name="toehold" class="nav">Toehold Switch</a></h2>
 
<h2><a name="toehold" class="nav">Toehold Switch</a></h2>
 +
    <p>Our experiments were performed with the D and G LacZ switches from the paper from the Collins group, <a href="http://www.cell.com/abstract/S0092-8674(14)01291-4" target="_blank">"Paper-Based Synthetic Gene Networks"</a>.</p>
 +
    <h3>RNA Trigger</h3>
 +
    <p>In <a href="https://2016.igem.org/Team:Pittsburgh/Notebook#Week5" target="_blank">Week 5</a>, we showed that LacZ is expressed when the plasmids for the switch and trigger constructs are combined in a cell-free system.</p>
 +
    <h3>DNA Trigger</h3>
 +
    <p>In <a href="https://2016.igem.org/Team:Pittsburgh/Notebook#Week6" target="_blank">Week 6</a>, we showed that toehold switches are activated by single-stranded DNA triggers, but activation levels are lower than with RNA triggers.</p>
 +
    <h3>Sequestered DNA Trigger</h3>
 +
    <p>In <a href="https://2016.igem.org/Team:Pittsburgh/Notebook#Week7" target="_blank">Week 7</a>, we showed that single-stranded DNA triggers do not activate toehold switches if they are part of a hybridized complex.</p>
 
     <a href="#top">Back to Top</a>
 
     <a href="#top">Back to Top</a>
 
<h2><a name="amplification" class="nav">Signal Amplification System</a></h2>  
 
<h2><a name="amplification" class="nav">Signal Amplification System</a></h2>  

Revision as of 18:49, 26 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

Contents

For our daily activities and experiments, please visit our Notebook. For a list of our protocols, please visit our Protocols page.

Reporter

amilCP

amilCP is a chromoprotein added to the iGEM Registry by Uppsala Sweden, 2012. We were unable to produce amilCP in bacteria or in our cell-free system.

Back to Top

Cell-Free System

Linear versus Plasmid DNA

We have good data from adding plasmids to our cell-free system. However, because the Collins paper specified the use of linear DNA in their cell-free systems, we compared the signal from GFP from linear and plasmid DNA. There is no siginificant difference between the two constructs.

dilution

volume optimization

Back to Top

DNAzyme

Back to Top

Toehold Switch

Our experiments were performed with the D and G LacZ switches from the paper from the Collins group, "Paper-Based Synthetic Gene Networks".

RNA Trigger

In Week 5, we showed that LacZ is expressed when the plasmids for the switch and trigger constructs are combined in a cell-free system.

DNA Trigger

In Week 6, we showed that toehold switches are activated by single-stranded DNA triggers, but activation levels are lower than with RNA triggers.

Sequestered DNA Trigger

In Week 7, we showed that single-stranded DNA triggers do not activate toehold switches if they are part of a hybridized complex.

Back to Top

Signal Amplification System

Back to Top

Differential Amplifier

Back to Top