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| | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Demonstrate">Demonstrate</a></li> | | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Demonstrate">Demonstrate</a></li> |
| | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Applied_Design">Applied design</a></li> | | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Applied_Design">Applied design</a></li> |
| | + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/InterLab">InterLab</a></li> |
| | + | |
| | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Notebook">Notebook</a></li> | | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Notebook">Notebook</a></li> |
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| | </li> | | </li> |
| | | | |
| − | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/InterLab">InterLab</a></li> | + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Medals">Medals</a></li> |
| | | | |
| | </ul> | | </ul> |
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| | <div class="div_nav"> | | <div class="div_nav"> |
| | <nav class="toc_nav" id="toc_show"> | | <nav class="toc_nav" id="toc_show"> |
| − | <ul> | + | |
| − | <li>
| + | |
| − | <a href="#Fluorescein">Fluorescein curve</a>
| + | |
| − | <ul>
| + | |
| − | <li><a href="#Fluorescein-Plate-reader">Plate reader</a></li>
| + | |
| − | <li><a href="#Fluorescein-Material">Material</a></li>
| + | |
| − | <li><a href="#Fluorescein-Method">Method</a></li>
| + | |
| − | <li><a href="#Fluorescein-Data-result">Data result</a></li>
| + | |
| − | </ul>
| + | |
| − | </li>
| + | |
| − | <li>
| + | |
| − | <a href="#OD600">OD600 Reference point</a>
| + | |
| − | <ul>
| + | |
| − | <li><a href="#OD600-Plate-reader">Plate reader</a></li>
| + | |
| − | <li><a href="#OD600-Material">Material</a></li>
| + | |
| − | <li><a href="#OD600-Method">Method</a></li>
| + | |
| − | <li><a href="#OD600-Data-result">Data result</a></li>
| + | |
| − | </ul>
| + | |
| − | </li>
| + | |
| − | | + | |
| − | <li>
| + | |
| − | <a href="#Cell">Cell measure</a>
| + | |
| − | <ul>
| + | |
| − | <li><a href="#Cell-Material">Material</a></li>
| + | |
| − | <li><a href="#Cell-Method">Method</a></li>
| + | |
| − | <li><a href="#Cell-Data-result">Data result</a></li>
| + | |
| − | </ul>
| + | |
| − | </li>
| + | |
| − | | + | |
| − | | + | |
| − | </ul>
| + | |
| | <svg class="toc-marker" width="200" height="200" xmlns="http://www.w3.org/2000/svg"> | | <svg class="toc-marker" width="200" height="200" xmlns="http://www.w3.org/2000/svg"> |
| | <path stroke="#00dbff" stroke-width="3" fill="transparent" stroke-dasharray="0, 0, 0, 1000" stroke-linecap="round" stroke-linejoin="round" transform="translate(-0.5, -0.5)" /> | | <path stroke="#00dbff" stroke-width="3" fill="transparent" stroke-dasharray="0, 0, 0, 1000" stroke-linecap="round" stroke-linejoin="round" transform="translate(-0.5, -0.5)" /> |
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| | <article class="contents_nav"> | | <article class="contents_nav"> |
| | <section> | | <section> |
| | + | <img src="https://static.igem.org/mediawiki/2017/6/6c/DEVICE_APP_DES.jpg" style="display:block; margin:auto;"><br/> |
| | + | <p> |
| | + | The WHO estimates that nearly all adults will have dental caries at some point. In Taiwan, the people suffer from dental caries is up to 99%. Dental caries are easily ignored by patients since they do not hurt until they become serious. However, you usually have to make an appointment with the dentist beforehand in Taiwan. In the worst situation, your tooth decay became so serious, that a tooth implantation is required, which is a painful and costly procedure. <br/> |
| | + | But with Carindex, the things will change. Carindex is a device designed to detect customer's dental caries risk by simply spitting your saliva into a drawer and put it into Carindex; it will tell you your risk of caries. What’s more, all data will be collected and accumulated to build a curve that help customer keep track of your oral health condition. We hope, in the near future, Carindex will decrease the dental caries rate significantly for every individual. |
| | | | |
| − |
| |
| − | <div id="Fluorescein">
| |
| − | <h2>Fluorescein Fluorescence standard curve</h2>
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| − |
| |
| − | </div>
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| − |
| |
| − | <div id="Fluorescein-Plate-reader">
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| − | <div class="aaa"></div>
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| − | <h3>Plate reader</h3>
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| − |
| |
| − | <p>
| |
| − | microplate reader FLUOstar Omega</br>
| |
| − | emission filter: 520 nm</br>
| |
| − | excitation filter: 485 nm
| |
| − | </p>
| |
| − |
| |
| − | </div>
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| − |
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| − | <div id="Fluorescein-Material">
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| − | <div class="aaa"></div>
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| − | <h3>Material</h3>
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| − | <p>
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| − | Fluorescein sodium salt</br>
| |
| − | 1xPBS</br>
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| − | Tissue culture testplate (black with flat bottom)
| |
| − | </p>
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| − |
| |
| − | </div>
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| − |
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| − | <div id="Fluorescein-Method">
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| − | <div class="aaa"></div>
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| − | <h3>Method</h3>
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| − |
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| − | <ol><li>Prepare fluorescein stock solution</li></ol>
| |
| − | <p>
| |
| − | 1. Spin down fluorescein stock tube to make sure pellet is at the bottom of tube.</br>
| |
| − | 2. Prepare 2x fluorescein stock solution (100 μM) by resuspending fluorescein in 1 mL of 1xPBS. </br>
| |
| − | 3. Dilute the 2x fluorescein stock solution with 1xPBS to make a 1x fluorescein solution and resulting concentration of fluorescein stock solution 50 μM </br></br>
| |
| − | </p>
| |
| − | <ol><li>Serial dilutions</li></ol>
| |
| − | <p>
| |
| − | 1. Add 100 μl of PBS into wells A2, B2, C2, D2....A12, B12, C12, D12</br>
| |
| − | 2. Add 200 μl of fluorescein 1x stock solution into A1, B1, C1, D1</br>
| |
| − | 3. Transfer 100 μl of fluorescein stock solution from A1 into A2. </br>
| |
| − | 4. Mix A2 by pipetting up and down 3x and transfer 100 μl into A3. </br>
| |
| − | 5. Mix A3 by pipetting up and down 3x and transfer 100 μl into A4. </br>
| |
| − | 6. Mix A4 by pipetting up and down 3x and transfer 100 μl into A5. </br>
| |
| − | 7. Mix A5 by pipetting up and down 3x and transfer 100 μl into A6. </br>
| |
| − | 8. Mix A6 by pipetting up and down 3x and transfer 100 μl into A7. </br>
| |
| − | 9. Mix A7 by pipetting up and down 3x and transfer 100 μl into A8. </br>
| |
| − | 10. Mix A8 by pipetting up and down 3x and transfer 100 μl into A9. </br>
| |
| − | 11. Mix A9 by pipetting up and down 3x and transfer 100 μl into A10.</br>
| |
| − | 12. Mix A10 by pipetting up and down 3x and transfer 100 μl into A11. </br>
| |
| − | 13. Mix A11 by pipetting up and down 3x and transfer 100 μl into liquid waste.
| |
| − | (Caution: Do not to continue serial dilution into column 12.)</br>
| |
| | </p> | | </p> |
| − | <ol><li>repeat serial dilute for Row B、D、E</strong></li></ol>
| |
| − | <ol><li>Measure fluorescence of all samples in all standard measurement modes in instrument Record the data in your notebook</strong></li></ol>
| |
| − | <ol><li>Import data into Excel (fluorescein standard curve tab ) Sheet_1 provided</li></ol>
| |
| − |
| |
| − | <br/><br/>
| |
| | | | |
| − | </div>
| |
| | | | |
| − | <div id="Fluorescein-Data-result">
| |
| − | <div class="aaa"></div>
| |
| − | <h3>Data result</h3>
| |
| − | <br/>
| |
| − | <img src="https://static.igem.org/mediawiki/2017/8/85/FFs_1.png" style="display:block; margin:auto;"><br/><br/>
| |
| − | <img src="https://static.igem.org/mediawiki/2017/a/a4/FFs_2.png" style="display:block; margin:auto;"><br/><br/>
| |
| − | <img src="https://static.igem.org/mediawiki/2017/5/5c/FFs_3.png" style="display:block; margin:auto;"><br/>
| |
| − | </div>
| |
| − | </section>
| |
| | | | |
| − | <section>
| |
| | | | |
| | | | |
| − | <div id="OD600">
| |
| − | <h2>OD600 Reference point</h2>
| |
| | | | |
| − | </div>
| |
| | | | |
| | | | |
| − | <div id="OD600-Plate-reader">
| |
| − | <div class="aaa"></div>
| |
| − | <h3>Plate reader</h3>
| |
| | | | |
| − | <p>
| |
| − | Thermo Scientific™ Multiskan™ FC Filter-based Microplate Photometer</br>
| |
| − | Filter: 595 nm</br>
| |
| − | </p>
| |
| | | | |
| − | </div>
| |
| | | | |
| − | <div id="OD600-Material">
| |
| − | <div class="aaa"></div>
| |
| − | <h3>Material</h3>
| |
| − | <p>
| |
| − | 1 ml LUDOX</br>
| |
| − | mQH<sub>2</sub>O</br>
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| − | 96 well cell culture plate (clear with flat-bottom)
| |
| − | </p>
| |
| | | | |
| − | </div>
| |
| | | | |
| − | <div id="OD600-Method">
| |
| − | <div class="aaa"></div>
| |
| − | <h3>Method</h3>
| |
| − |
| |
| − | <p>
| |
| − | 1. Add 100 μl LUDOX into wells A1, B1, C1, D1 (or 1 mL LUDOX into cuvette)</br>
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| − | 2. Add 100 μl of H<sub>2</sub>O into wells A2, B2, C2, D2 (or 1 mL H<sub>2</sub>O into cuvette)</br>
| |
| − | 3. Measure absorbance 600 nm of all samples in all standard measurement modes in instrument</br>
| |
| − | 4. Record the data in excel and Import data into Excel ( OD600 reference point tab ) Sheet_1 provided</br>
| |
| − | </p>
| |
| − |
| |
| − | </div>
| |
| − |
| |
| − | <div id="OD600-Data-result">
| |
| − | <div class="aaa"></div>
| |
| − | <h3>Data result</h3>
| |
| − | <br/>
| |
| − | <img src="https://static.igem.org/mediawiki/2017/c/cd/Ee.jpeg" style="display:block; margin:auto;"><br/>
| |
| − | </div>
| |
| − | </section>
| |
| − |
| |
| − | <section>
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| − |
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| − |
| |
| − | <div id="Cell">
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| − | <h2>Cell measure</h2>
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| − |
| |
| − | </div>
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| − |
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| − |
| |
| − | <div id="Cell-Material">
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| − | <div class="aaa"></div>
| |
| − | <h3>Material</h3>
| |
| − | <p>
| |
| − | Competent cells ( Escherichia coli strain DH5α)</br>
| |
| − | LB (Luria Bertani) media</br>
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| − | Chloramphenicol (stock concentration 25 mg/mL dissolved in EtOH - working stock 25 μg /mL)</br>
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| − | 50 ml Falcon tube (or equivalent, preferably amber or covered in foil to block light)</br>
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| − | Incubator at 37°C</br>
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| − | 1.5 ml eppendorf tubes for sample storage</br>
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| − | Ice bucket with ice</br>
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| − | Pipettes</br>
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| − | 96 well plate(cell culture 96 well plate、tissue culture testplate)</br>
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| − | Devices (from InterLab Measurement Kit):</br>
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| − | 1. Negative control(BBa_R0040)</br>
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| − | 2. Positive control(J23151+B0032+E0040+B0010+B0012)</br>
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| − | 3. Test Device 1: J23101+I13504</br>
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| − | 4. Test Device 2: J23106+I13504</br>
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| − | 5. Test Device 3: J23117+I13504</br>
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| − | 6. Test Device 4: J23101+BCD2+E0040+B0015</br>
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| − | 7. Test Device 5: J23106+BCD2+E0040+B0015</br>
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| − | 8. Test Device 6: J23117+BCD2+E0040+B0015</br>
| |
| − | </p>
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| − |
| |
| − | </div>
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| − |
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| − | <div id="Cell-Method">
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| − | <div class="aaa"></div>
| |
| − | <h3>Method</h3>
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| − |
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| − | <p>
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| − | 1. Day 1 : Resuspended each plasmid in plate 7 and transform into Escherichia coli DH5α.</br>
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| − | (Transformation protocol is from iGEM)</br>
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| − | 2. Day 2 : Pick 2 colonies from each of plate and inoculate it on 5-10 mL LB medium +Chloramphenicol.Grow the cells overnight (16-18 hours) at 37°C and 170 rpm.</br>
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| − | 3. Day 3 : Set instrument to read OD600 (as OD calibration setting)and measure OD600 of the overnight cultures</br>
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| − | 4. Dilute the cultures to a target OD 600 of 0.02 in 12 ml LB medium + Chloramphenicol in 50 mL falcon tube (covered with foil to block light).</br>
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| − | 5. Incubate the cultures at 37°C and 170 rpm.</br>
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| − | 6. Take 1000 μL samples of the cultures at 0, 2, 4, and 6 hours of incubation and place samples on ice</br>
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| − | 7. 4 replicates of 100 uL samples were taken from each culture at 0, 2, 4, and 6 hours of incubation and placed in a 96 well plate for OD and fluorescence measurements using the setup described above</br>
| |
| − | </p>
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| − |
| |
| − | </div>
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| − |
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| − | <div id="Cell-Data-result">
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| − | <div class="aaa"></div>
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| − | <h3>Data result</h3>
| |
| − | <br/>
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| − | <img src="https://static.igem.org/mediawiki/2017/f/f7/IL_cell1.jpg" style="display:block; margin:auto;"><br/><br/>
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| − | <img src="https://static.igem.org/mediawiki/2017/4/41/IL_cell2.jpg" style="display:block; margin:auto;"><br/>
| |
| − | </div>
| |
| | </section> | | </section> |
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| | <!-- Contact --> | | <!-- Contact --> |
| − | <section class="contact"> | + | |
| − |
| + | |
| − | <ul class="icons">
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| − | <li>
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| − | <a href="https://www.facebook.com/ccuigemteam" target="_blank">
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| − | <img src="https://static.igem.org/mediawiki/2016/2/2f/T--Harvard_BioDesign--images_facebook01.png"alt="Facebook Logo" style="width:51px;height:51px;">
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| − | </a>
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| − | <a href="emailto:ccu.igem.2017@gmail.com">
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| − | <img src="https://static.igem.org/mediawiki/2016/e/e2/T--Harvard_BioDesign--images_gmail01.png" alt="Email Logo" style="width:51px;height:51px;">
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| − | </ul>
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| − | </section>
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| | | | |
| | <!-- Copyright --> | | <!-- Copyright --> |
| | + | <!-- Copyright --> |
| | <div class="copyright"> | | <div class="copyright"> |
| − | <ul class="menu"> | + | |
| − | <li>© 2017 CCU Taiwan iGEM</li><li>Design: <a href="http://html5up.net">HTML5 UP</a></li>
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| − | </ul>
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| | </div> | | </div> |
| − | | + | <img src="https://static.igem.org/mediawiki/2017/f/fb/Ccu.jpg" width="400" height="120" style="margin-top: 5em; |
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| | </div> | | </div> |
| | </div> | | </div> |
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| | </div> | | </div> |
| | + | |
| | + | <!-- Scripts --> |
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| | </body> | | </body> |
| | </html> | | </html> |
