Difference between revisions of "Team:Bielefeld-CeBiTec/Results/it-really-works"
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<div class="jumbotron" style="background-image: url(https://static.igem.org/mediawiki/2015/2/2b/Bielefeld-CeBiTec_CFPS_image_paper_discs_general.png)"> | <div class="jumbotron" style="background-image: url(https://static.igem.org/mediawiki/2015/2/2b/Bielefeld-CeBiTec_CFPS_image_paper_discs_general.png)"> | ||
<div class="jumbotron-text"> | <div class="jumbotron-text"> | ||
| − | <h1 style="margin-bottom: 0px"> | + | <h1 style="margin-bottom: 0px">All-in-One Performance</h1> |
<p>It LITERALLY works on paper!</p> | <p>It LITERALLY works on paper!</p> | ||
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<h2> Project aim: Achieved! </h2> | <h2> Project aim: Achieved! </h2> | ||
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<figure style="width: 350px; margin-right: 20px; float: left"> | <figure style="width: 350px; margin-right: 20px; float: left"> | ||
<a href="https://static.igem.org/mediawiki/2015/0/08/Bielefeld-CeBiTec_prototype_box_and_smartphone_up_big.png" data-lightbox="finalaim" data-title="Don't drink this water!"><img src="https://static.igem.org/mediawiki/2015/7/71/Bielefeld-CeBiTec_prototype_box_and_smartphone_up_small.png" alt="box"></a> | <a href="https://static.igem.org/mediawiki/2015/0/08/Bielefeld-CeBiTec_prototype_box_and_smartphone_up_big.png" data-lightbox="finalaim" data-title="Don't drink this water!"><img src="https://static.igem.org/mediawiki/2015/7/71/Bielefeld-CeBiTec_prototype_box_and_smartphone_up_small.png" alt="box"></a> | ||
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<a href="https://static.igem.org/mediawiki/2015/f/f6/Bielefeld-CeBiTec_all_in_one_box_seite_big.png" data-lightbox="finalaim" data-title="Prototype box for detection"><img src="https://static.igem.org/mediawiki/2015/c/c3/Bielefeld-CeBiTec_all_in_one_box_seite_small.png" alt="Device box"></a> | <a href="https://static.igem.org/mediawiki/2015/f/f6/Bielefeld-CeBiTec_all_in_one_box_seite_big.png" data-lightbox="finalaim" data-title="Prototype box for detection"><img src="https://static.igem.org/mediawiki/2015/c/c3/Bielefeld-CeBiTec_all_in_one_box_seite_small.png" alt="Device box"></a> | ||
| − | <figcaption> Our measurement prototype for easy and fast evaluation of | + | <figcaption> Our measurement prototype for easy and fast evaluation of fluorescence emission </figcaption> |
</figure> | </figure> | ||
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| + | <p>We aimed at developing a cell-free test strip for the detection of various substances at once. We started with many different subteams. Some people cloned the different biosensors, others were occupied with the ideal design of the teststrip, the development of a cell-free detection approach or the processing of the output signal. So in our last weeks our main focus was to bring all this different aspects together to one working approach. And we did it <b>successfully</b>! </p> | ||
| + | <p> We achieved <b>a lot</b> during this project, just take a look at this (far from complete) list to get an idea:</p> | ||
<ol> | <ol> | ||
| − | <li> Highly efficient and robust cell-free synthesis of sfGFP <b>on simple paper</b> that can be stored</li> | + | <li> Highly efficient and robust cell-free synthesis of sfGFP <b>on simple paper</b> that can be stored. By the way, you just have to spend <b>16 ¢</b> for each reaction</li> |
<li> Cloning and characterization of <b>multiple</b> heavy metal sensors</li> | <li> Cloning and characterization of <b>multiple</b> heavy metal sensors</li> | ||
<li> Cloning and characterization of a <b>date rape drug</b> sensor </li> | <li> Cloning and characterization of a <b>date rape drug</b> sensor </li> | ||
<li> Establishment of a <b>smartphone based fluorescence detection method</b></li> | <li> Establishment of a <b>smartphone based fluorescence detection method</b></li> | ||
| − | <li> Implementing of an app to easily evaluate the measured fluorescence signals.</li> | + | <li> Implementing of an <b>app</b> to easily evaluate the measured fluorescence signals.</li> |
</ol> | </ol> | ||
</br> | </br> | ||
| − | + | <p> So this is, how they actually work together! </p> | |
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<ol> | <ol> | ||
| − | <li> At first, you have to combine all reagents that are necessary to conduct cell-free protein synthesis | + | <li> At first, you have to combine all reagents that are necessary to conduct cell-free protein synthesis. </li> |
<li> When all compounds are combined, the reaction is applied on paper and freeze-dried. This procedure prevents the reaction from premature starting. Additionally, all remaining cells are killed.</li> | <li> When all compounds are combined, the reaction is applied on paper and freeze-dried. This procedure prevents the reaction from premature starting. Additionally, all remaining cells are killed.</li> | ||
<li> The paper with the reaction is storable and transportable. For examination of water quality, you can simply add a water drop to the paper. </li> | <li> The paper with the reaction is storable and transportable. For examination of water quality, you can simply add a water drop to the paper. </li> | ||
| − | <li> In no time you can take a picture of the paper with your smartphone. A filter combination enables you to capture the fluorescence that is emitted by the reporter protein </li> | + | <li> In no time you can take a picture of the paper with your smartphone. A filter combination for your smartphone enables you to capture the fluorescence that is emitted by the reporter protein. </li> |
<li> Our app evaluates the data and tells you if your water sample is contaminated. </li> | <li> Our app evaluates the data and tells you if your water sample is contaminated. </li> | ||
</ol> | </ol> | ||
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</div> | </div> | ||
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| − | <p> In this way, we made it possible to detect various heavy metals. We proved this by using mercury contaminated water for rehydration of two different setups: One was our standard cell-free reaction setup and the other was our mercury detector. We took a picture, and our app | + | |
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| + | <video muted controls src = "https://static.igem.org/mediawiki/2015/8/84/Bielefeld-CeBiTec_AllinOneVideo.mp4" type="video/mp4" width="1060" | ||
| + | height="525" | ||
| + | poster="https://static.igem.org/mediawiki/2015/0/08/Bielefeld-CeBiTec_prototype_box_and_smartphone_up_big.png"> | ||
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| + | Let´s hope your browser supports this! But if you can read this, that´s probably not the case. | ||
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| + | </video> | ||
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| + | <p> In this way, we made it possible to detect various heavy metals. We proved this by using mercury and copper contaminated water for rehydration of two different setups: One was our standard cell-free reaction setup and the other was our mercury detector. We took a picture, and our app immediately gave us the result: <b>"Your water is contaminated with: Mercury and Copper"!</b></p> | ||
<div class="row"> | <div class="row"> | ||
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<figure style="width: 450px; margin-right: 20px; float: left"> | <figure style="width: 450px; margin-right: 20px; float: left"> | ||
<a href="https://static.igem.org/mediawiki/2015/e/e8/Bielefeld-CeBiTec_all_in_one_hg_without_filter_big.png" data-lightbox="finalaim" data-title="Cell-free biosensor for mercury detection in water samples, pictured without filters."><img src="https://static.igem.org/mediawiki/2015/6/66/Bielefeld-CeBiTec_all_in_one_hg_without_filter_small.png" alt="Cell free Hg sensor"></a> | <a href="https://static.igem.org/mediawiki/2015/e/e8/Bielefeld-CeBiTec_all_in_one_hg_without_filter_big.png" data-lightbox="finalaim" data-title="Cell-free biosensor for mercury detection in water samples, pictured without filters."><img src="https://static.igem.org/mediawiki/2015/6/66/Bielefeld-CeBiTec_all_in_one_hg_without_filter_small.png" alt="Cell free Hg sensor"></a> | ||
| − | <figcaption> Paper discs | + | <figcaption> Paper discs containing the cell-free standards and Hg sensor, pictured without filters. </figcaption> |
</figure> | </figure> | ||
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</div> | </div> | ||
</div> | </div> | ||
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| + | <p> We consider our cell-free biosensor an outstanding instrument for the detection of harmful substances in liquids. </p> | ||
| + | <p> We see great need for a storable, transportable and easy-to-use water quality test strip all around the world.</p> | ||
| + | <p> Our development can help to improve the quality of living by revealing the quality of water. </p> | ||
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| − | <p> During the reaction we had measured the fluorescence in a plate reader. In the nearby figures you can see: The standard <a data-toggle="tooltip" title="Cell-Free Protein Synthesis">CFPS</a> setup performed | + | <p> During the reaction we had measured the fluorescence in a plate reader. In the nearby figures you can see: The standard <a data-toggle="tooltip" title="Cell-Free Protein Synthesis">CFPS</a> setup performed not sufficent when mercury was present, HOWEVER, when using our mercury detection setup, cell-free synthesis of our reporter protein sfGFP was greatly enhanced! </p> |
<figure style="width: 500px; margin-right: 20px; float: left"> | <figure style="width: 500px; margin-right: 20px; float: left"> | ||
Revision as of 02:42, 19 September 2015
All-in-One Performance
It LITERALLY works on paper!
Project aim: Achieved!
We aimed at developing a cell-free test strip for the detection of various substances at once. We started with many different subteams. Some people cloned the different biosensors, others were occupied with the ideal design of the teststrip, the development of a cell-free detection approach or the processing of the output signal. So in our last weeks our main focus was to bring all this different aspects together to one working approach. And we did it successfully!
We achieved a lot during this project, just take a look at this (far from complete) list to get an idea:
- Highly efficient and robust cell-free synthesis of sfGFP on simple paper that can be stored. By the way, you just have to spend 16 ¢ for each reaction
- Cloning and characterization of multiple heavy metal sensors
- Cloning and characterization of a date rape drug sensor
- Establishment of a smartphone based fluorescence detection method
- Implementing of an app to easily evaluate the measured fluorescence signals.
So this is, how they actually work together!
- At first, you have to combine all reagents that are necessary to conduct cell-free protein synthesis.
- When all compounds are combined, the reaction is applied on paper and freeze-dried. This procedure prevents the reaction from premature starting. Additionally, all remaining cells are killed.
- The paper with the reaction is storable and transportable. For examination of water quality, you can simply add a water drop to the paper.
- In no time you can take a picture of the paper with your smartphone. A filter combination for your smartphone enables you to capture the fluorescence that is emitted by the reporter protein.
- Our app evaluates the data and tells you if your water sample is contaminated.
In this way, we made it possible to detect various heavy metals. We proved this by using mercury and copper contaminated water for rehydration of two different setups: One was our standard cell-free reaction setup and the other was our mercury detector. We took a picture, and our app immediately gave us the result: "Your water is contaminated with: Mercury and Copper"!
We consider our cell-free biosensor an outstanding instrument for the detection of harmful substances in liquids.
We see great need for a storable, transportable and easy-to-use water quality test strip all around the world.
Our development can help to improve the quality of living by revealing the quality of water.
During the reaction we had measured the fluorescence in a plate reader. In the nearby figures you can see: The standard CFPS setup performed not sufficent when mercury was present, HOWEVER, when using our mercury detection setup, cell-free synthesis of our reporter protein sfGFP was greatly enhanced!
At molecular level, our biosensors work on basis of one simple concept: The expression of a gene is regulated by a protein. In case of a transcriptional repressor, the reporter protein is not expressed unless a particular analyte is present. In case of a transcriptional activator, expression of the reporter protein is enhanced in presence of the analyte.
This regulation is a central process in nature. Because our system uses this concept, it is highly extensible and can be used to tackle various real world problems.
We realized that date rape drug intoxications are currently one of these problems. With our development we introduce an easy and unprecedented date rape drug ingredient biosensor! To detect the date rape drug ingredient γ-hydroxybutyrate (GHB), we had to adjust only two components of our setup: The repressor protein and the DNA sequence that the protein recognizes.
