Difference between revisions of "Team:Valencia UPV/Demonstrate"

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                  ">Index</h4>
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                        ">Index</h4>
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                          ">Printeria device concept-testing</a>
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                                ">Printeria device concept-testing</a>
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                          ">Printeria Wet Lab demonstrations</a>
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                                ">Printeria Wet Lab demonstrations</a>
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            <a class="anchorOffset" id="Demonstrate"></a><h2 class="h2Demonstrate">Demonstrate</h2><p>
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                  <a class="anchorOffset" id="Demonstrate"></a><h2 class="h2Demonstrate">Demonstrate</h2><p>
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              After long months of effort and dedication, the <b>Printeria project of the Valencia UPV iGEM 2018 team is now a reality!</b> All our work has been reflected in our Printeria device. With this project we intend to make the approach to Synthetic Biology a reality, but... to what extent has our team been able to develop Printeria? Does it really work? <b>Let's see all the evidences of our achievements...</b>
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                    After long months of effort and dedication, the <b>Printeria project of the Valencia UPV iGEM 2018 team is now a reality!</b> All our work has been reflected in our Printeria device. With this project we intend to make the approach to Synthetic Biology a reality, but... to what extent has our team been able to develop Printeria? Does it really work? <b>Let's see all the evidences of our achievements...</b>
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              </p><a class="anchorOffset" id="concept-testing"></a><h4>Printeria device concept-testing</h4><p>
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                    </p><a class="anchorOffset" id="concept-testing"></a><h4>Printeria device concept-testing</h4><p>
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              <b>Here we show you all the evidence to convince you that the Printeria device really works...</b>
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                    <b>Here we show you all the evidence to convince you that the Printeria device really works...</b>
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          <li><p>
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          The <b>input system</b> is a completely original approach in order to dispense precise quantities of liquid in a compact and highly controllable way. You can check in this video how our device input system works...
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                        The <a href="https://2018.igem.org/Team:Valencia_UPV/Hardware#Entry" target="_blank"><b>input system</b></a> is a completely original approach in order to dispense precise quantities of liquid in a compact and highly controllable way. You can check in this video how our device input system works...
  
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          VIDEO
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          The <b>digital MicroFluidics</b> is at the centre of Printeria, and it enables a high <a href="" target="_blank">control of the reactions</a>. We have accomplished the movement on the droplets in smaller test boards, as you can see here:
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                      <a href="https://static.igem.org/mediawiki/2018/d/db/T--Valencia_UPV--ENTRADA5UPV2018.gif" data-lightbox="true">
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          And then we moved to a more clean product that is able to hold a larger number of pads and it is easier to control. You can see that result below:
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                      <h6 style="text-align: left; padding-left: 5em;">Proof of concept of the input system.</h6>
  
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          VIDEOS
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                        The <b>digital MicroFluidics</b> is at the centre of Printeria, and it enables a high <a href="https://2018.igem.org/Team:Valencia_UPV/Hardware#Reaction" target="_blank">control of the reactions</a>. We have accomplished the movement on the droplets in smaller test boards, as you can see here:
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          The droplet needs to be heated and cooled for the reaction to take place. We have implemented these <b>hot and cold zones</b> via a <a href="" target="_blank">resistor and a Peltier</a>. Here the stability of the temperature control for both is demonstrated.
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          GRÁFICA
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          We have included the design of out <b>electroporator</b> from <a href="https://2016.igem.org/Team:Valencia_UPV" target="_blank" style="padding-right: 0">Hype It iGEM project</a>, developed in 2016 by Valencia UPV team. The results of our experiment using it are shown here:
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          GRÁFICA
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          For the output we have included an <b>Optical Density and Fluorescence sensors</b> in order to measure data from the bacteria that are growing on the <a href="" target="_blank">tube</a>. The results of our tests are the following:
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                        And then we moved to a more clean product that is able to hold a larger number of pads and it is easier to control. You can see that result below: 
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</p><h6 style="text-align: left; padding-left: 5em;">OD measurements registred by Printeria OD sensor adn fitted to an exponential curve of cell growth.</h6>
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GRAFICA FLUORESCENCIA
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              </ul><a class="anchorOffset" id="Wetlab_dem"></a><h4>Printeria Wet Lab demonstrations</h4><p>
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                      VIDEOS
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              <b>In this section you can check all our Wet Lab demonstrations...</b>
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                        The droplet needs to be heated and cooled for the reaction to take place. We have implemented these <b>hot and cold zones</b> via a resistor and two peltiers. Here the stability of the temperature control for both is demonstrated.
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                      GRÁFICA
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                        We have included the design of out <b>electroporator</b> from <a href="https://2016.igem.org/Team:Valencia_UPV" target="_blank" style="padding-right: 0">Hype It iGEM project</a>, developed in 2016 by Valencia UPV team. The results of our experiment using it are shown here:
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              We have proved that assembling functional composite parts with the <a href="" target="_blank"> Golden Gate method</a> with our basic parts is possible... Here you can see a <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2656103" target="_blank">sfGFP transcriptional unit</a> that has been obtained by employing Golden Gate assembly method.
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                      GRÁFICA
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                        For the output we have included an <b>Optical Density and Fluorescence sensors</b> in order to measure data from the bacteria that are growing on the <a href="https://2018.igem.org/Team:Valencia_UPV/Hardware#Measurements" target="_blank">tube</a>. The results of our tests are the following:
  
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                        </p><h6 style="text-align: left; padding-left: 5em;">OD measurements registred by Printeria OD sensor adn fitted to an exponential curve of cell growth.</h6>
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                        GRAFICA FLUORESCENCIA
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<h6 style="text-align: left; padding-left: 5em;">Assembly of BBa_K2656103 transformed into electrocompetent bacteria.</h6>
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                    </ul><a class="anchorOffset" id="Wetlab_dem"></a><h4>Printeria Wet Lab demonstrations</h4><p>
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                    <b>In this section you can check all our Wet Lab demonstrations...</b>
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              It has also been proven that by using <b>linearized destination vectors</b> in the Golden Gate assembly reactions, a very small number of bacteria transformed with vectors without the desired insert are obtained. Here you can see a <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2656105" target="_blank">GFP transcriptional unit</a> obtained by using a linearized destination vector in the assembly reaction.
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                          We have proved that assembling functional composite parts with the <a href="" target="_blank"> Golden Gate method</a> with our basic parts is possible... Here you can see a <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2656103" target="_blank">sfGFP transcriptional unit</a> that has been obtained by employing Golden Gate assembly method.
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              Another demonstration is that electrocompetent bacteria can be stored at -20ºC for at least two weeks and chilled for 4 hours at room temperature (30.3ºC) without losing their competence.
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              </p><a href="https://static.igem.org/mediawiki/2018/e/ea/T--Valencia_UPV--competent_cellsUPV2018.png" data-lightbox="true">
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margin-right: 0px !important;text-align: left; padding-left: 0em;">Serial decimal dilutions (from left to right: 10<sup>-2</sup>, 10<sup>-1</sup> and 10<sup>0</sup>) of transformed electrocompetent bacteria stored at -20°C for two weeks and set at Room Temperature (37°C) 4 hours before electroporation. These bacteria are transformed with P10500 and plated in LB-agar petri dishes with cloranphenicol. </h6></div><div style="
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            Finally, to compare the efficiency of <b>Golden Gate vs BioBricks assembly</b>, we have designed a <a href="" target="_blank">comparative experiment</a> between two TU of identical structure but assembled with Golden Gate and BioBricks method, respectively. Our results demonstrate that both methods have the same efficiency.
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            </p></div><a href="https://static.igem.org/mediawiki/2018/1/17/T--Valencia_UPV--comparison_GGBB_graphUPV2018.png" data-lightbox="true">
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</a><h6 style="text-align: left; padding-left: 5em;">Comparison between fluorescence and absorbance of Golden Gate and BioBrick TU.</h6><div>
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                <h6 style="text-align: left; padding-left: 5em;">Assembly of BBa_K2656103 transformed into electrocompetent bacteria.</h6>
  
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                          It has also been proven that by using <b>linearized destination vectors</b> in the Golden Gate assembly reactions, a very small number of bacteria transformed with vectors without the desired insert are obtained. Here you can see a <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2656105" target="_blank">GFP transcriptional unit</a> obtained by using a linearized destination vector in the assembly reaction.
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                          Another demonstration is that electrocompetent bacteria can be stored at -20ºC for at least two weeks and chilled for 4 hours at room temperature (30.3ºC) without losing their competence.
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                        </p><a href="https://static.igem.org/mediawiki/2018/e/ea/T--Valencia_UPV--competent_cellsUPV2018.png" data-lightbox="true">
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                <img src="https://static.igem.org/mediawiki/2018/e/ea/T--Valencia_UPV--competent_cellsUPV2018.png">
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      margin-right: 0px !important;text-align: left; padding-left: 0em;">Serial decimal dilutions (from left to right: 10<sup>-2</sup>, 10<sup>-1</sup> and 10<sup>0</sup>) of transformed electrocompetent bacteria stored at -20°C for two weeks and set at Room Temperature (37°C) 4 hours before electroporation. These bacteria are transformed with P10500 and plated in LB-agar petri dishes with cloranphenicol. </h6></div><div style="
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                        Finally, to compare the efficiency of <b>Golden Gate vs BioBricks assembly</b>, we have designed a <a href="" target="_blank">comparative experiment</a> between two TU of identical structure but assembled with Golden Gate and BioBricks method, respectively. Our results demonstrate that both methods have the same efficiency.
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               One of the Printeria Wetlab challenges during the project has been to demonstrate that Printeria transcriptional units (TU) obtained with <a href="" target="_blank">Golden Gate assembly technology</a> are as efficients as TU assembled by BioBrick technology. The main advantage offered by Golden Gate technology is the posibility to perform an assembly reaction with many parts in a single step. This is the main reason why we have design an extensive <a href="https://2018.igem.org/Team:Valencia_UPV/Part_Collection" target="_blank">Part Collection</a> standarized to GoldenBraid 3.0 grammar. However, <b>which is the efficiency of these parts compared to BioBrick parts?</b>
 
               One of the Printeria Wetlab challenges during the project has been to demonstrate that Printeria transcriptional units (TU) obtained with <a href="" target="_blank">Golden Gate assembly technology</a> are as efficients as TU assembled by BioBrick technology. The main advantage offered by Golden Gate technology is the posibility to perform an assembly reaction with many parts in a single step. This is the main reason why we have design an extensive <a href="https://2018.igem.org/Team:Valencia_UPV/Part_Collection" target="_blank">Part Collection</a> standarized to GoldenBraid 3.0 grammar. However, <b>which is the efficiency of these parts compared to BioBrick parts?</b>
 
               In this way we can confirm, thanks to the results obtained, that <b>the efficiency will be the same independently of the assembly method</b>.
 
               In this way we can confirm, thanks to the results obtained, that <b>the efficiency will be the same independently of the assembly method</b>.
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               <b>For the correct working of Printeria many technologies have to interact and work together</b> in order to produce the desired reaction. But the individual parts themselves also have to work properly in order for the whole to function. In this way, we want to <b>our individual proofs of concept</b> that then are joined on Printeria so that finally our user does not have to think about the complex processes that happen inside Printeria.
 
               <b>For the correct working of Printeria many technologies have to interact and work together</b> in order to produce the desired reaction. But the individual parts themselves also have to work properly in order for the whole to function. In this way, we want to <b>our individual proofs of concept</b> that then are joined on Printeria so that finally our user does not have to think about the complex processes that happen inside Printeria.
 
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               The <b>Input system</b> is a completely original approach in order to dispense precise quantities of liquid in a compact and highly controllable way. As well as to make it easier for the user to introduce the materials all at once or individually (discover more about the inner workings of <a href="" target="_blank">Printeria input system</a>). You can see our concept test on this video:
 
               The <b>Input system</b> is a completely original approach in order to dispense precise quantities of liquid in a compact and highly controllable way. As well as to make it easier for the user to introduce the materials all at once or individually (discover more about the inner workings of <a href="" target="_blank">Printeria input system</a>). You can see our concept test on this video:
  
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The <b>digital MicroFluidics</b> is at the centre of Printeria, and it enables a high <a href="" target="_blank">control of the reactions</a>. We have accomplished the movement on the droplets in smaller test boards, as you can see here:
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        The <b>digital MicroFluidics</b> is at the centre of Printeria, and it enables a high <a href="" target="_blank">control of the reactions</a>. We have accomplished the movement on the droplets in smaller test boards, as you can see here:
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          The droplet needs to be heated and cooled for the reaction to take place. We have implemented these <b>hot and cold zones</b> via a <a href="" target="_blank">resistor and a Peltier</a>. Here the stability of the temperature control for both is demonstrated.
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                  The droplet needs to be heated and cooled for the reaction to take place. We have implemented these <b>hot and cold zones</b> via a <a href="" target="_blank">resistor and a Peltier</a>. Here the stability of the temperature control for both is demonstrated.
  
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Revision as of 14:44, 11 October 2018

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Index

Demonstrate

After long months of effort and dedication, the Printeria project of the Valencia UPV iGEM 2018 team is now a reality! All our work has been reflected in our Printeria device. With this project we intend to make the approach to Synthetic Biology a reality, but... to what extent has our team been able to develop Printeria? Does it really work? Let's see all the evidences of our achievements...

Printeria device concept-testing

Here we show you all the evidence to convince you that the Printeria device really works...

  • The input system is a completely original approach in order to dispense precise quantities of liquid in a compact and highly controllable way. You can check in this video how our device input system works...

    • Proof of concept of the input system.

  • The digital MicroFluidics is at the centre of Printeria, and it enables a high control of the reactions. We have accomplished the movement on the droplets in smaller test boards, as you can see here:

    And then we moved to a more clean product that is able to hold a larger number of pads and it is easier to control. You can see that result below:

    • VIDEOS

  • The droplet needs to be heated and cooled for the reaction to take place. We have implemented these hot and cold zones via a resistor and two peltiers. Here the stability of the temperature control for both is demonstrated.

    • GRÁFICA

  • We have included the design of out electroporator from Hype It iGEM project, developed in 2016 by Valencia UPV team. The results of our experiment using it are shown here:

    • GRÁFICA

  • For the output we have included an Optical Density and Fluorescence sensors in order to measure data from the bacteria that are growing on the tube. The results of our tests are the following:

    OD measurements registred by Printeria OD sensor adn fitted to an exponential curve of cell growth.
    GRAFICA FLUORESCENCIA

Printeria Wet Lab demonstrations

In this section you can check all our Wet Lab demonstrations...

We have proved that assembling functional composite parts with the Golden Gate method with our basic parts is possible... Here you can see a sfGFP transcriptional unit that has been obtained by employing Golden Gate assembly method.

Assembly of BBa_K2656103 transformed into electrocompetent bacteria.

It has also been proven that by using linearized destination vectors in the Golden Gate assembly reactions, a very small number of bacteria transformed with vectors without the desired insert are obtained. Here you can see a GFP transcriptional unit obtained by using a linearized destination vector in the assembly reaction.

Transformed bacteria resulting from the assembly of BBa_K2656105 TU.

Another demonstration is that electrocompetent bacteria can be stored at -20ºC for at least two weeks and chilled for 4 hours at room temperature (30.3ºC) without losing their competence.

Serial decimal dilutions (from left to right: 10-2, 10-1 and 100) of transformed electrocompetent bacteria stored at -20°C for two weeks and set at Room Temperature (37°C) 4 hours before electroporation. These bacteria are transformed with P10500 and plated in LB-agar petri dishes with cloranphenicol.

Finally, to compare the efficiency of Golden Gate vs BioBricks assembly, we have designed a comparative experiment between two TU of identical structure but assembled with Golden Gate and BioBricks method, respectively. Our results demonstrate that both methods have the same efficiency.

Comparison between fluorescence and absorbance of Golden Gate and BioBrick TU.
POSIBILIDAD DE HACER UN TEST ESTADÍSTICO O DAR ALGUN DATO QUE PRUEBE QUE SON IGUALES FALTAN AÑADIR REFERENCIAS HARDWARE Y EXPERIMENTOS

CONTACT US igem.upv.2018@gmail.com