Difference between revisions of "Team:Munich/Demonstrate"

Line 104: Line 104:
 
<tr><td class="verticalColumn" colspan=2 align=center valign=center>
 
<tr><td class="verticalColumn" colspan=2 align=center valign=center>
 
<h3>Sensitive and Rapid</h3>
 
<h3>Sensitive and Rapid</h3>
<!-- <img width=300> -->
+
<img width=300 src="https://static.igem.org/mediawiki/2017/6/6e/T--Munich--Demonstrate_image1.png">
 
</td>
 
</td>
 
<td colspan=2 align=center valign=center>
 
<td colspan=2 align=center valign=center>
 
<h3>Robust and Universal</h3>  
 
<h3>Robust and Universal</h3>  
<!-- <img width=300> -->
+
<img width=300 src="https://static.igem.org/mediawiki/2017/9/94/T--Munich--Demonstrate_image2.png">
 
</td>
 
</td>
 
<td colspan=2 align=center valign=center>
 
<td colspan=2 align=center valign=center>
 
<h3>Portable and low-cost</h3>
 
<h3>Portable and low-cost</h3>
<!-- <img width=300-->
+
<img width=300 src="https://static.igem.org/mediawiki/2017/3/3b/T--Munich--Demonstrate_image3.png">
 
</td>
 
</td>
 
</tr>
 
</tr>
  
 
<tr><td colspan=6 align=center valign=center>
 
<tr><td colspan=6 align=center valign=center>
<img src="https://static.igem.org/mediawiki/2017/a/a5/T--Munich--Demonstrate_Graph_Descriptions.svg">
 
 
<h3>Sample processing</h3>
 
<h3>Sample processing</h3>
 
<p>
 
<p>

Revision as of 23:07, 1 November 2017


Demonstrate

With our modular approach, we prototype all units of our diagnostic device, and integrate them into a customizable platform. We use cell-free synthetic biology to distinguish between pathogens with our universal detection cascade.

Sensitive and Rapid

Robust and Universal

Portable and low-cost

Sample processing

We chose to combine heat lysis and isothermal amplification (RPA) to extract our target RNA from patient samples.

  • We used RPA to amplify DNA from heat lysed E. coli.
  • We conducted RPA and transcription from an in vitro DNA on paper.
  • We amplified and transcribed an in vitro DNA target to RNA concentrations detectable by our readout circuit .

We validated the three necessary modules of the sample processing (cell lysis, DNA amplification and transcription) with a rapid and sensitive method, RPA.

Readout circuit

We chose Cas13a for pathogen identification because of its specificity for nucleic acid sequence detection.

  • We characterized Cas13a and its detection limit with native and lyophilized protein, with in vitro and in vivo sources of RNA, in bulk and on paper.
  • We designed fluorescent and colorimetric readouts, and used a synthetic aptamer as a detection tool.
  • We tested targets from common pathogens and showed the orthogonality of virus detection versus bacterial detection.

We proved the robustness and universality of our Cas13a-based fluorescence readout circuit.

Readout circuit

We chose a disposable paper strip combined with a reusable fluorescence detector to analyse our samples.

  • We optimized the paper support for Cas13a functionality.
  • We built the most sensitive and cheapest fluorescence detector ever created by an iGEM team to our knowledge.
  • We successfully used our fluorescence detector to detect Cas13a activity on paper.

We created a detection chip that is portable, functional and affordable, for the distribution of our diagnosis device, CascAID.