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| − | <meta charset="utf-8"><link rel="shortcut icon" href="/wiki/images/8/80/T--NCKU_Tainan--favicon.png" type="image/x-icon"><link rel="icon" type="image/png" href="/wiki/images/8/80/T--NCKU_Tainan--favicon.png"><link rel="icon" type="image/x-icon" href="/wiki/images/8/80/T--NCKU_Tainan--favicon.png"><meta name="viewport" content="width=device-width, initial-scale=1.0"><meta property="og:title" content="Notebook Functional Test - iGEM NCKU"><meta property="og:site_name" content="Notebook Functional Test - iGEM NCKU"><meta property="og:description" content=""><title>Notebook Functional Test - iGEM NCKU</title><meta http-equiv="Content-Type" content="text/html" charset="utf-8"><meta property="og:image" content=""><meta property="og:image:type" content="image/png"><link rel="stylesheet" href="/Team:NCKU_Tainan/css/frame/T--NCKU_Tainan--bootstrap_min_css?ctype=text/css&action=raw"><link href="/Team:NCKU_Tainan/font/T--NCKU_Tainan--NotoSans_css?ctype=text/css&action=raw" rel="stylesheet" type="text/css"><link rel="stylesheet" href="/Team:NCKU_Tainan/font/T--NCKU_Tainan--font-awesome_min_css?ctype=text/css&action=raw"> <link rel="stylesheet" href="/Team:NCKU_Tainan/css/T--NCKU_Tainan--Notebook_css?ctype=text/css&action=raw"> | + | <meta charset="utf-8"><link rel="shortcut icon" href="/wiki/images/8/80/T--NCKU_Tainan--favicon.png" type="image/x-icon"><link rel="icon" type="image/png" href="/wiki/images/8/80/T--NCKU_Tainan--favicon.png"><link rel="icon" type="image/x-icon" href="/wiki/images/8/80/T--NCKU_Tainan--favicon.png"><meta name="viewport" content="width=device-width, initial-scale=1.0"><meta property="og:title" content="Notebook Functional Test - iGEM NCKU"><meta property="og:site_name" content="Notebook Functional Test - iGEM NCKU"><meta property="og:description" content=""><title>Notebook Functional Test - iGEM NCKU</title><meta http-equiv="Content-Type" content="text/html" charset="utf-8"><meta property="og:image" content=""><meta property="og:image:type" content="image/png"><link rel="stylesheet" href="/Team:NCKU_Tainan/css/frame/T--NCKU_Tainan--bootstrap_min_css?ctype=text/css&action=raw"><link href="/Team:NCKU_Tainan/font/T--NCKU_Tainan--NotoSans_css?ctype=text/css&action=raw" rel="stylesheet" type="text/css"><link rel="stylesheet" href="/Team:NCKU_Tainan/font/T--NCKU_Tainan--font-awesome_min_css?ctype=text/css&action=raw"> <link rel="stylesheet" href="/Team:NCKU_Tainan/css/T--NCKU_Tainan--Notebook_css?ctype=text/css&action=raw"> |
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| − | <style>@font-face { font-family: 'NotoSansCJKtc-Regular'; src: url("/wiki/images/0/0b/T--NCKU_Tainan--NotoSansCJKtc-Regular.woff") format('woff');}</style><nav class="navbar navbar-default"><div class=" container- fluid" style=" width:72%"> <div class="navbar-header"> <button type="button" class="navbar-toggle collapsed" data-toggle="collapse" data-target="#navbar" aria-expanded="false"> <span class="sr-only">Toggle navigation</span> <span class="icon-bar"></span> <span class="icon-bar"></span> <span class="icon-bar"></span> </button> <a class="navbar-brand" href="/Team:NCKU_Tainan"> <h1>NCKU</h1><h4>Tainan</h4> </a> </div> <div id="navbar" class="navbar-collapse collapse"> <ul class="nav navbar-nav"> <li> <a class="dropdown-toggle" type="button" id="dropdownMenu1" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="">Project</a><div class="nav-underline"></div> <ul class="dropdown-menu" aria-labelledby="dropdownMenu2"> <li><a href="/Team:NCKU_Tainan/Project"> Overview</a></li> <li><a href="/Team:NCKU_Tainan/Description">Description</a></li> <li><a href="/Team:NCKU_Tainan/Results">Results</a></li> <li><a href="/Team:NCKU_Tainan/Model">Modeling</a></li> <li><a href="/Team:NCKU_Tainan/Parts">Parts</a></li> </ul> </li> <li> <a class="dropdown-toggle" type="button" id="dropdownMenu2" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="">Device</a><div class="nav-underline"></div> <ul class="dropdown-menu" aria-labelledby="dropdownMenu2"> <li><a href="/Team:NCKU_Tainan/Hardware">Hardware</a></li> <li><a href="/Team:NCKU_Tainan/Software">Software</a></li> <li><a href="/Team:NCKU_Tainan/Demonstrate">Demonstrate</a></li> </ul> </li> <li> <a class="dropdown-toggle" type="button" id="dropdownMenu3" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="">Judging</a><div class="nav-underline"></div> <ul class="dropdown-menu" aria-labelledby="dropdownMenu3"> <li><a href="/Team:NCKU_Tainan/Medal">Medal</a></li> <li><a href="/Team:NCKU_Tainan/Safety">Safety</a></li> </ul> </li> <li> <a class="dropdown-toggle" type="button" id="dropdownMenu4" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="/Team:NCKU_Tainan/Team">Team</a><div class="nav-underline"></div> <ul class="dropdown-menu" aria-labelledby="dropdownMenu4"> <li><a href="/Team:NCKU_Tainan/Team">Team</a></li> <li><a href="/Team:NCKU_Tainan/Attributions">Attributions</a></li> <li><a href="/Team:NCKU_Tainan/Acknowledgement">Acknowledgement</a></li> <li><a href="/Team:NCKU_Tainan/Collaborations">Collaborations</a></li> </ul> </li> <li> <a class="dropdown-toggle" type="button" id="dropdownMenu5" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="/Team:NCKU_Tainan/Human_Practices">Human Practices</a><div class="nav-underline"></div> <ul class="dropdown-menu" aria-labelledby="dropdownMenu5"> <li><a href="/Team:NCKU_Tainan/Human_Practices">Overview</a></li> <li><a href="/Team:NCKU_Tainan/Integrated_Practices">Integrated Practices</a></li> <li><a href="/Team:NCKU_Tainan/Engagement">Engagement</a></li> </ul> </li> <li> <a class="dropdown-toggle" type="button" id="dropdownMenu6" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="">Notebook</a><div class="nav-underline"></div> <ul class="dropdown-menu" aria-labelledby="dropdownMenu6"> <li><a href="/Team:NCKU_Tainan/Notebook_Construction">Construction</a></li> <li><a href="/Team:NCKU_Tainan/Notebook_Functional_Test">Functional Test</a></li> <li><a href="/Team:NCKU_Tainan/Notebook_Device_Design">Device Design</a></li> <li><a href="/Team:NCKU_Tainan/Notebook_Protocols">Protocols</a></li> </ul> </li> </ul> <ul class="nav navbar-nav navbar-right"> </ul> </div><!--/.nav-collapse --></div><!--/.container-fluid --></nav><div id="container-big"><div id="iGEMbar"></div><div id="line-left"></div><div id="line-left2"></div><div id="line-right"></div><div id="photo-left"></div></div><!--/.container-big --> <style> | + | <style>@font-face { font-family: 'NotoSansCJKtc-Regular'; 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| − | #photo-left { background-image: url("/wiki/images/c/c1/T--NCKU_Tainan--sample2.jpg"); }
| + | #photo-left { background-image: url("/wiki/images/c/cb/T--NCKU_Tainan--Functional.png"); } |
| − | </style>
| + | </style> |
| − | <div class="container-fluid" style="margin-top:100px"> | + | <div class="container-fluid" style="margin-top:100px"> |
| − | <div class="head">NOTE / Functional Test</div> | + | <div class="head">NOTE / Functional Test</div> |
| − | <div class="content row"> | + | <div class="content row"> |
| − | | + | |
| − | <div class="head2">Notebook - Functional Test</div> | + | <div class="head2">Notebook - Functional Test</div> |
| − | <div class="title-line" id="sec1"></div> | + | <div class="title-line long" id="sec1"></div> |
| − | <div class="title-content"></div> | + | <div class="title-content"> Compare promoter efficiency of Pcar with PI</div> |
| − | <h5></h5>
| + | <div class="text-content"> |
| − | | + | <h5>May 14/2016</h5> |
| − | <h5></h5>
| + | <p> Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-Pcar-RBS-amilCP-TT and pMD19T-PI-RBS-amilCP-TT from LB agar plate.</p> |
| − | | + | <h5>May 15/2016</h5> |
| − | <h5></h5>
| + | <p> Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p> |
| − | | + | <h5>May 16/2016</h5> |
| − | | + | <p> Induction: add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration: 0, 5, 15, 30, 60, 120 mM</p> |
| − | <h5></h5>
| + | <p> Readouts: measure OD 580 kinetically for 3.5 hr and compare the efficiency of Pcar and PI</p> |
| − | | + | </div> |
| − | | + | <div class="title-line long" id="sec2"></div> |
| | + | <div class="title-content">Glucose-Color detection test</div> |
| | + | <div class="text-content"> |
| | + | <h5>May 20/2016</h5> |
| | + | <p> Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-amilCP-TT from LB agar plate.</p> |
| | + | <h5>May 21/2016</h5> |
| | + | <p> Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p > |
| | + | <p>Induction: add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration: 0, 5, 15, 30, 60, 120 mM.</p> |
| | + | <p>Readouts: measure OD 580 kinetically for 3.5 hr.</p> |
| | + | </div> |
| | | | |
| − | </div>
| + | <div class="title-line long" id="sec3"></div> |
| − | | + | <div class=" title- content">Urine Glucose- Color detection test</div> |
| − | <ul id="sidemenu">
| + | <div class="text-content"> |
| − | <li><a href="#" onclick="toEvent('section1');">Section 1</a></li>
| + | <h5>Jun. 17/2016</h5> |
| − | <li><a href="#" onclick="toEvent('section2');">Section 2</a></li>
| + | <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-amilCP-TT from LB agar plate.</p> |
| − | </ul>
| + | <h5>Jun. 18/2016</h5> |
| − | </div>
| + | <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p> |
| − | </div>
| + | <p>Sample preparation: collect urine sample from non-diabetic healthy individuals, and add additional glucose to final urine glucose concentration of 0, 10, 30, 120, 240 mM.</p> |
| − | </div> <!-- /.container-fluid -->
| + | <p>Induction: add 100 μl of bacteria culture to 96 well plate and add Sample Urine to final concentration: 0, 5, 15, 30, 60, 120 mM.</p> |
| − | <script src="/Team:NCKU_Tainan/js/frame/T--NCKU_Tainan--jquery-1_12_0_min_js?ctype=text/javascript&action=raw"></script><script src="/Team:NCKU_Tainan/js/frame/T--NCKU_Tainan--bootstrap_min_js?ctype=text/javascript&action=raw"></script><script src="/Team:NCKU_Tainan/js/T--NCKU_Tainan--MathjaxConfigIgem_js?ctype=text/javascript&action=raw"></script><script src="/common/MathJax-2.5-latest/MathJax.js?config=TeX-AMS-MML_HTMLorMML"></script><script src="/Team:NCKU_Tainan/js/T--NCKU_Tainan--common_js?ctype=text/javascript&action=raw"></script><script>(function() { /* change icon */ var link = document.createElement('link'); link.type = 'image/x-icon'; link.rel = 'shortcut icon'; link.href = '/wiki/images/8/80/T--NCKU_Tainan--favicon.png'; document.getElementsByTagName('head')[0].appendChild(link);}());</script>
| + | <p>Readouts: measure OD 580 kinetically for 3.5 hr.</p> |
| | + | </div> |
| | + | |
| | + | <div class="title-line long" id="sec4"></div> |
| | + | <div class="title-content">2-hour Glucose-Fluoresence detection test</div> |
| | + | <div class="text-content"> |
| | + | <h5>Jul. 15/2016</h5> |
| | + | <p>Transformed plasmid (pMD19T-PpI-B0034-E1010) to BL21 competent cell.</p> |
| | + | <h5>Jul. 16/2016</h5> |
| | + | <p>Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one colony from the agar plate.</p> |
| | + | <h5>Jul. 17/2016</h5> |
| | + | <p>Extracted plasmid and confirmed plasmid size by gel electrophoresis.<br> |
| | + | <i class="glyphicon glyphicon-ok"></i>Result: The size of pMD19T-PI-RBS-mRFP was correct.</p> |
| | + | <p>Culture: 10 ml of LB (with Ampicillin) cultured with 100 μl pre-cultured bacteria.</p> |
| | + | <h5>Jul. 18/2016</h5> |
| | + | <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP from LB agar plate.</p> |
| | + | <h5>Jul. 19/2016</h5> |
| | + | <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p> |
| | + | <p>Induction: add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration: 0, 5, 15, 30, 60, 120 mM.</p> |
| | + | <p>Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 2 hr.</p> |
| | + | </div> |
| | + | |
| | + | <div class="title-line long" id="sec5"></div> |
| | + | <div class="title-content">12-hour Glucose-Fluoresence detection test</div> |
| | + | <div class="text-content"> |
| | + | <h5>Jul. 25/2016</h5> |
| | + | <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP-TT from LB agar plate.</p> |
| | + | <h5>Jul. 26/2016</h5> |
| | + | <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p> |
| | + | <p>Induction: add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration: 0, 5, 15, 30, 60, 120 mM.</p> |
| | + | <p>Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 12 hr.</p> |
| | + | </div> |
| | + | |
| | + | <div class="title-line long" id="sec6"></div> |
| | + | <div class="title-content">Lysis test</div> |
| | + | <div class="text-content"> |
| | + | <h5>Aug. 22/2016</h5> |
| | + | <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-P<sub>BAD</sub>-RBS-lysis-TT from LB agar plate.</p> |
| | + | <h5>Aug. 23/2016</h5> |
| | + | <p>Transfer: 20 ml of modified M9 (with Ampicilin) added with different glucose/arabinose concentration and culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 15 mins and transfer to 96 well plate.</p> |
| | + | <p>Readouts: measure OD 600 kinetically with plate readerfor 4 hours.</p> |
| | + | </div> |
| | + | |
| | + | <div class="title-line long" id="sec7"></div> |
| | + | <div class="title-content">Lysis test-2</div> |
| | + | <div class="text-content"> |
| | + | <h5>Sep. 01/2016</h5> |
| | + | <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-P<sub>BAD</sub>-RBS-lysis-TT from LB agar plate.</p> |
| | + | <h5>Sep. 02/2016</h5> |
| | + | <p>Transfer: 20 ml of modified M9 (with Ampicilin)added with different glucose/arabinose concentration and culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 15 mins and transfer to 96 well plate.</p> |
| | + | <p>Readouts: measure OD 600 kinetically with plate readerfor 4 hours.</p> |
| | + | </div> |
| | + | |
| | + | <div class="title-line long" id="sec8"></div> |
| | + | <div class="title-content">2-hour Urine Glucose-Fluoresence detection test</div> |
| | + | <div class="text-content"> |
| | + | <h5>Sep. 05/2016</h5> |
| | + | <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP-TT from LB agar plate.</p> |
| | + | <h5>Sep. 06/2016</h5> |
| | + | <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p> |
| | + | <p>Sample preparation: collect urine sample from non-diabetic healthy individuals, and add additional glucose to final urine glucose concentration of 0, 10, 30, 120, 240 mM.</p> |
| | + | <p>Induction: add 100 ul of bacteria culture to 96 well plate and add urine sample to final concentration: 0, 5, 15, 30, 60, 120 mM</p> |
| | + | <p>Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 2 hr.</p> |
| | + | </div> |
| | + | |
| | + | <div class="title-line long" id="sec9"></div> |
| | + | <div class="title-content">12-hour Glucose-Fluoresence detection test-1 </div> |
| | + | <div class="text-content"> |
| | + | <h5>Sep. 12/2016</h5> |
| | + | <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP-TT from LB agar plate.</p> |
| | + | <h5>Sep. 13/2016</h5> |
| | + | <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p> |
| | + | <p>Sample preparation: collect urine sample from non-diabetic healthy individuals, and add additional glucose to final urine glucose concentration of 0, 10, 30, 120, 240 mM.</p> |
| | + | <p>Induction: add 100 μl of bacteria culture to 96 well plate and add urine sample to final concentration: 0, 5, 15, 30, 60, 120 mM.</p> |
| | + | <p>Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 12 hr.</p> |
| | + | </div> |
| | + | </div> |
| | + | </div> |
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