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− | {{CCU_Taiwan}}
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− | <h1>Notebook</h1> | + | <script type="text/javascript" src="https://2017.igem.org/Team:CCU_Taiwan/assets/js/js_anchor? |
− | <p> Document the dates you worked on your project. This should be a detailed account of the work done each day for your project.</p> | + | action=raw&ctype=text/javascript"></script> |
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| + | </head> |
| + | <body class="no-sidebar" class="body.homepage"> |
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| + | <div class="fixed" style="cursor:pointer;"> |
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| </div> | | </div> |
− | <div class="clear"></div>
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| + | <div id="page-wrapper"> |
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− | <div class="column half_size"> | + | <!-- Header --> |
− | <h5>What should this page have?</h5> | + | <div id="header"> |
− | <ul> | + | |
− | <li>Chronological notes of what your team is doing.</li> | + | <!-- Inner --> |
− | <li> Brief descriptions of daily important events.</li> | + | <div class="inner"> |
− | <li>Pictures of your progress. </li> | + | <header> |
− | <li>Mention who participated in what task.</li> | + | <h1><a href="https://2017.igem.org/Team:CCU_Taiwan" id="logo">Attributions</a></h1> |
− | </ul> | + | |
| + | |
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| + | |
| + | </header> |
| + | </div> |
| + | |
| + | <!-- Nav --> |
| + | <nav id="nav"> |
| + | <ul> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan">Home</a></li> |
| + | |
| + | |
| + | <li> |
| + | <a href="#">Project</a> |
| + | <ul> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Description">Description</a></li> |
| + | <li> |
| + | <a href="https://2017.igem.org/Team:CCU_Taiwan/Design">Biosensor</a> |
| + | <ul> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Design#CSP">CSP detector</a></li> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Design#Lactate">Lactate detector</a></li> |
| + | </ul> |
| + | </li> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Test_paper">Test paper</a></li> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Experiments">Experiments</a></li> |
| + | |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Results">Results</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/Notebook">Notebook</a></li> |
| + | |
| + | </ul> |
| + | </li> |
| + | |
| + | <li> |
| + | |
| + | <a href="#">Hardware</a> |
| + | <ul> |
| + | |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Hardware">Hardware overview</a></li> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Device_design">Device design</a></li> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Device_detection">Device detection</a></li> |
| + | </ul> |
| + | </li> |
| + | |
| + | <li> |
| + | <a href="#">Software</a> |
| + | <ul> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/IOT system">IOT system</a></li> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/APP">APP</a></li> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Machine_learning">Machine learning</a></li> |
| + | </ul> |
| + | </li> |
| + | |
| + | </li> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Model">Modeling</a></li> |
| + | |
| + | <li> |
| + | <a href="#">Human practice</a> |
| + | <ul> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Human_Practices">Human practice overview</a></li> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/HP/Silver">Silver HP</a></li> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/HP/Gold_Integrated">Integrate & Gold</a></li> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Engagement">Public engagemant</a></li> |
| + | |
| + | </ul> |
| + | </li> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Entrepreneurship">Entrepreneurship</a></li> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Safety">Safety</a></li> |
| + | <li> |
| + | <a href="#">Parts</a> |
| + | <ul> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Parts">Parts</a></li> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Basic_Part">Basic parts</a></li> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Composite_Part">Composite parts</a></li> |
| + | |
| + | </ul> |
| + | </li> |
| + | |
| + | |
| + | |
| + | <li> |
| + | <a href="#">Team</a> |
| + | <ul> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Members">Team members</a></li> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Attributions">Attributions</a></li> |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/Collaborations">Collaborations</a></li> |
| + | |
| + | </ul> |
| + | </li> |
| + | |
| + | <li><a href="https://2017.igem.org/Team:CCU_Taiwan/InterLab">InterLab</a></li> |
| + | |
| + | </ul> |
| + | </nav> |
| + | |
| + | </div> |
| + | |
| + | <!-- Main --> |
| + | <div class="wrapper style1"> |
| + | |
| + | <div class="container"> |
| + | <article id="main" class="special"> |
| + | <header> |
| + | |
| + | |
| + | <div class="div_nav"> |
| + | <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"> |
| + | <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)" /> |
| + | </svg> |
| + | </nav> |
| + | |
| + | <article class="contents_nav"> |
| + | <section> |
| + | |
| + | |
| + | <div id="Fluorescein"> |
| + | <h2>Fluorescein Fluorescence standard curve</h2> |
| | | |
| </div> | | </div> |
| | | |
− | <div class="column half_size"> | + | <div id="Fluorescein-Plate-reader"> |
− | <h5>Inspiration</h5> | + | <div class="aaa"></div> |
− | <p>You can see what others teams have done to organize their notes:</p> | + | <h3>Plate reader</h3> |
| | | |
− | <ul> | + | <p> |
− | <li><a href="https://2014.igem.org/Team:ATOMS-Turkiye/Notebook">2014 ATOMS-Turkiye</a></li> | + | microplate reader FLUOstar Omega</br> |
− | <li><a href="https://2014.igem.org/Team:Tec-Monterrey/ITESM14_project.html#tab_notebook">2014 Tec Monterrey</a></li>
| + | emission filter: 520 nm</br> |
− | <li><a href="https://2014.igem.org/Team:Kyoto/Notebook/Magnetosome_Formation#title">2014 Kyoto</a></li>
| + | excitation filter: 485 nm |
− | <li><a href="https://2014.igem.org/Team:Cornell/notebook">2014 Cornell</a></li>
| + | </p> |
− | </ul> | + | |
| | | |
| </div> | | </div> |
| + | |
| + | <div id="Fluorescein-Material"> |
| + | <div class="aaa"></div> |
| + | <h3>Material</h3> |
| + | <p> |
| + | Fluorescein sodium salt</br> |
| + | 1xPBS</br> |
| + | Tissue culture testplate (black with flat bottom) |
| + | </p> |
| + | |
| + | </div> |
| + | |
| + | <div id="Fluorescein-Method"> |
| + | <div class="aaa"></div> |
| + | <h3>Method</h3> |
| + | |
| + | <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> |
| + | <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> |
| + | 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> |
| + | 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> |
| + | |
| + | |
| + | <div id="Cell"> |
| + | <h2>Cell measure</h2> |
| + | |
| + | </div> |
| + | |
| + | |
| + | <div id="Cell-Material"> |
| + | <div class="aaa"></div> |
| + | <h3>Material</h3> |
| + | <p> |
| + | Competent cells ( Escherichia coli strain DH5α)</br> |
| + | LB (Luria Bertani) media</br> |
| + | Chloramphenicol (stock concentration 25 mg/mL dissolved in EtOH - working stock 25 μg /mL)</br> |
| + | 50 ml Falcon tube (or equivalent, preferably amber or covered in foil to block light)</br> |
| + | Incubator at 37°C</br> |
| + | 1.5 ml eppendorf tubes for sample storage</br> |
| + | Ice bucket with ice</br> |
| + | Pipettes</br> |
| + | 96 well plate(cell culture 96 well plate、tissue culture testplate)</br> |
| + | Devices (from InterLab Measurement Kit):</br> |
| + | 1. Negative control(BBa_R0040)</br> |
| + | 2. Positive control(J23151+B0032+E0040+B0010+B0012)</br> |
| + | 3. Test Device 1: J23101+I13504</br> |
| + | 4. Test Device 2: J23106+I13504</br> |
| + | 5. Test Device 3: J23117+I13504</br> |
| + | 6. Test Device 4: J23101+BCD2+E0040+B0015</br> |
| + | 7. Test Device 5: J23106+BCD2+E0040+B0015</br> |
| + | 8. Test Device 6: J23117+BCD2+E0040+B0015</br> |
| + | </p> |
| + | |
| + | </div> |
| + | |
| + | <div id="Cell-Method"> |
| + | <div class="aaa"></div> |
| + | <h3>Method</h3> |
| + | |
| + | <p> |
| + | 1. Day 1 : Resuspended each plasmid in plate 7 and transform into Escherichia coli DH5α.</br> |
| + | (Transformation protocol is from iGEM)</br> |
| + | 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> |
| + | 3. Day 3 : Set instrument to read OD600 (as OD calibration setting)and measure OD600 of the overnight cultures</br> |
| + | 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> |
| + | 5. Incubate the cultures at 37°C and 170 rpm.</br> |
| + | 6. Take 1000 μL samples of the cultures at 0, 2, 4, and 6 hours of incubation and place samples on ice</br> |
| + | 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> |
| + | |
| + | </div> |
| + | |
| + | <div id="Cell-Data-result"> |
| + | <div class="aaa"></div> |
| + | <h3>Data result</h3> |
| + | <br/> |
| + | <img src="https://static.igem.org/mediawiki/2017/f/f7/IL_cell1.jpg" style="display:block; margin:auto;"><br/><br/> |
| + | <img src="https://static.igem.org/mediawiki/2017/4/41/IL_cell2.jpg" style="display:block; margin:auto;"><br/> |
| + | </div> |
| + | </section> |
| + | |
| + | </article> |
| + | </div> |
| + | |
| + | |
| + | |
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| + | </div> |
| + | |
| + | </div> |
| + | |
| + | |
| + | <!-- Footer --> |
| + | <div id="footer"> |
| + | <div class="container"> |
| + | <div class="row"> |
| + | <div class="12u"> |
| + | |
| + | <!-- Contact --> |
| + | <section class="contact"> |
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| + | <ul class="icons"> |
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| + | <li> |
| + | <a href="https://www.facebook.com/ccuigemteam" target="_blank"> |
| + | <img src="https://static.igem.org/mediawiki/2016/2/2f/T--Harvard_BioDesign--images_facebook01.png"alt="Facebook Logo" style="width:51px;height:51px;"> |
| + | </a> |
| + | </li> |
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| + | <a href="emailto:ccu.igem.2017@gmail.com"> |
| + | <img src="https://static.igem.org/mediawiki/2016/e/e2/T--Harvard_BioDesign--images_gmail01.png" alt="Email Logo" style="width:51px;height:51px;"> |
| + | </a> |
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| + | </ul> |
| + | </section> |
| + | |
| + | <!-- 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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| + | </div> |
| + | </div> |
| + | </div> |
| + | |
| + | </div> |
| + | |
| + | |
| + | </body> |
| </html> | | </html> |