Difference between revisions of "Team:CCU Taiwan/HP/Gold Integrated"

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<head>
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<h3>★  ALERT! </h3>
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<title>No Sidebar - Helios by HTML5 UP</title>
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<p>This page is used by the judges to evaluate your team for the <a href="https://2017.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2017.igem.org/Judging/Awards"> award listed above</a>. </p>
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<p> Delete this box in order to be evaluated for this medal criterion and/or award. See more information at <a href="https://2017.igem.org/Judging/Pages_for_Awards"> Instructions for Pages for awards</a>.</p>
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<body class="no-sidebar">
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<div id="page-wrapper">
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<!-- Header -->
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<div id="header">
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<!-- Inner -->
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<div class="inner">
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<header>
 +
<h1><a href="https://2017.igem.org/Team:CCU_Taiwan" id="logo">Gold Integrated</a></h1>
 +
 
 +
 
 +
 
 +
 
 +
 
 +
 
 +
</header>
 +
</div>
 +
 
 +
<!-- Nav -->
 +
<nav id="nav">
 +
<ul>
 +
 +
 
 +
 
 +
<li>
 +
<a href="#">Project</a>
 +
<ul>
 +
<li><a href="#">Description</a></li>
 +
                                                                                <li>
 +
<a href="#">Biosensor</a>
 +
<ul>
 +
<li><a href="#">CSP detector</a></li>
 +
<li><a href="#">Lactate detector</a></li>
 +
</ul>
 +
</li>
 +
<li><a href="#">Modeling</a></li>
 +
<li><a href="#">Experiment</a></li>
 +
 +
<li><a href="Results">Result</a></li>
 +
<li><a href="#">Future perspective</a></li>
 +
<li><a href="#">Safty</a></li>
 +
</ul>
 +
</li>
 +
 
 +
<li>
 +
<a href="#">Dry lab</a>
 +
<ul>
 +
                                                                                <li>
 +
<a href="#">Hardware</a>
 +
<ul>
 +
<li><a href="#">Device</a></li>
 +
<li><a href="#">Detector</a></li>
 +
</ul>
 +
</li>
 +
 
 +
                                                                                <li>
 +
<a href="#">Software</a>
 +
<ul>
 +
<li><a href="#">IOT system</a></li>
 +
<li><a href="#">APP</a></li>
 +
                                                                                                <li><a href="#">Machine learning</a></li>
 +
</ul>
 +
</li>
 +
</ul>
 +
</li>
 +
 
 +
 
 +
<li>
 +
<a href="#">Human practice</a>
 +
<ul>
 +
<li><a href="#">Silver</a></li>
 +
<li><a href="#">Gold and Integrated</a></li>
 +
<li><a href="#">Engagemant</a></li>
 +
<li><a href="#">Entrepreneurship</a></li>
 +
</ul>
 +
</li>
 +
 
 +
<li>
 +
<a href="#">Part</a>
 +
<ul>
 +
<li><a href="#">Basic part</a></li>
 +
<li><a href="#">Composite part</a></li>
 +
 +
</ul>
 +
</li>
 +
 
 +
 
 +
<li>
 +
<a href="#">Notebook</a>
 +
<ul>
 +
<li><a href="#">Schedule</a></li>
 +
<li><a href="#">Protocol</a></li>
 +
<li><a href="https://2017.igem.org/Team:CCU_Taiwan/InterLab">InterLab</a></li>
 +
 +
</ul>
 +
</li>
 +
 
 +
 
 +
<li>
 +
<a href="#">Team</a>
 +
<ul>
 +
<li><a href="https://2017.igem.org/Team:CCU_Taiwan/Member">Member</a></li>
 +
<li><a href="https://2017.igem.org/Team:CCU_Taiwan/Collaborations">Collaborations</a></li>
 +
<li><a href="#">Attribution</a></li>
 +
 +
</ul>
 +
</li>
 +
 
 +
 
 +
<li>
 +
<a href="#">Judging</a>
 +
<ul>
 +
<li><a href="#">Gold</a></li>
 +
<li><a href="#">Silver</a></li>
 +
<li><a href="#">Bronze</a></li>
 +
 +
</ul>
 +
</li>
 +
 
 +
 
 +
 
 +
 
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<li><a href="no-sidebar.html">Q&A</a></li>
 +
</ul>
 +
</nav>
 +
 
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</div>
 +
 
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<!-- Main -->
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<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="#444" 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 class="aaa"></div>
 
</div>
 
</div>
−
<div class="clear"></div>
 
  
−
<div class="column full_size">
+
<div id="Fluorescein-Plate-reader">
  
−
<h1>Gold Medal and Integrated Human Practices</h1>
+
<h3>Plate reader</h3>
  
−
<p>This page will contain information for your Gold medal Human Practices work, which you can also use to nominate your team for the Best Integrated Human Practices page. To make things easier, we have combined the Gold medal page with the Best Integrated Human Practices page since we expect the work to overlap considerably. </p>
+
    <p>
−
<p>iGEM teams are unique and leading the field because they "go beyond the lab" to imagine their projects in a social/environmental context, to better understand issues that might influence the design and use of their technologies.</p>
+
      microplate reader FLUOstar Omega</br>
−
<p>Teams work with students and advisors from the humanities and social sciences to explore topics concerning ethical, legal, social, economic, safety or security issues related to their work. Consideration of these Human Practices is crucial for building safe and sustainable projects that serve the public interest. </p>
+
emission filter: 520 nm</br>
−
<p>For more information, please see the <a href="https://2017.igem.org/Competition/Human_Practices">Human Practices page</a>.</p>
+
excitation filter: 485 nm
 +
    </p>
 +
<div class="aaa"></div>
 
</div>
 
</div>
−
<div class="clear"></div>
+
 
 +
<div id="Fluorescein-Material">
  
 +
<h3>Material</h3>
 +
<p>
 +
Fluorescein sodium salt</br>
 +
1xPBS</br>
 +
Tissue culture testplate (black with flat bottom)
 +
</p>
 +
<div class="aaa"></div>
 +
</div>
  
−
<div class="column half_size">
+
<div id="Fluorescein-Method">
−
<h3>Gold Medal Criterion #1</h3>
+
−
<p>Expand on your silver medal activity by demonstrating how you have integrated the investigated issues into the design and/or execution of your project.</p>
+
  
 +
                <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 class="aaa"></div>
 
</div>
 
</div>
 +
<div id="Fluorescein-Data-result">
  
−
<div class="column half_size">
+
<h3>Data result</h3>
−
<h3>Best Integrated Human Practices Special Prize</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 class="aaa"></div>
 +
</div>
 +
 
 +
 
 +
<div id="OD600-Plate-reader">
 +
 
 +
<h3>Plate reader</h3>
 +
 
 +
    <p>
 +
Thermo Scientific™ Multiskan™ FC Filter-based Microplate Photometer</br>
 +
Filter: 595 nm</br>
 +
    </p>
 +
<div class="aaa"></div>
 +
</div>
 +
 
 +
<div id="OD600-Material">
 +
 
 +
<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 class="aaa"></div>
 +
</div>
 +
 
 +
<div id="OD600-Method">
 +
 
 +
                <h3>Method</h3>
  
 
<p>
 
<p>
−
To compete for the <a href="https://2017.igem.org/Judging/Awards">Best Integrated Human Practices prize</a>, please describe your work on this page and also fill out the description on the <a href="https://2017.igem.org/Judging/Judging_Form">judging form</a>.
+
1. Add 100 μl LUDOX into wells A1, B1, C1, D1 (or 1 mL LUDOX into cuvette)</br>
−
<br><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>
−
You must also delete the message box on the top of this page to be eligible for this prize.
+
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>
 
</p>
 +
<div class="aaa"></div>
 
</div>
 
</div>
−
<div class="clear"></div>
+
 
−
<div class="column full_size">
+
<div id="OD600-Data-result">
−
<h5>Inspiration</h5>
+
 
−
<p>Here are a few examples of excellent Integrated Human Practices work:</p>
+
<h3>Data result</h3>
−
<ul>
+
<br/>
−
<li><a href="https://2016.igem.org/Team:INSA-Lyon/Integrated_Practices">2016 INSA Lyon</a></li>
+
<img src="https://static.igem.org/mediawiki/2017/5/5a/600_1.png" style="display:block; margin:auto;"><br/>
−
<li><a href="https://2016.igem.org/Team:UofC_Calgary/Integrated_Practices">2016 UofC Calgary</a></li>
+
</div>
−
<li><a href="https://2015.igem.org/Team:Bielefeld-CeBiTec/Practices">2015 Bielefeld</a></li>
+
</section>
−
<li><a href="https://2015.igem.org/Team:Edinburgh/Practices">2015 Edinburgh</a></li>
+
 
−
</ul>
+
<section>
 +
 
 +
 
 +
<div id="Cell">
 +
<h2>Cell measure</h2>
 +
<div class="aaa"></div>
 +
</div>
 +
 
 +
 
 +
<div id="Cell-Material">
 +
 
 +
<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 class="aaa"></div>
 +
</div>
 +
 
 +
<div id="Cell-Method">
 +
 
 +
                <h3>Method</h3>
 +
 
 +
<p>
 +
1. Day 1 : Resuspended each plasmid in plate 7 and transform into Escherichia coli DH5α.</br>
 +
&nbsp;(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 class="aaa"></div>
 +
</div>
 +
 
 +
<div id="Cell-Data-result">
 +
 
 +
<h3>Data result</h3>
 +
<br/>
 +
<img src="https://static.igem.org/mediawiki/2017/3/30/Cell_1.png" style="display:block; margin:auto;"><br/><br/>
 +
<img src="https://static.igem.org/mediawiki/2017/f/fe/Cell_2.png" style="display:block; margin:auto;"><br/>
 +
</div>
 +
</section>
 +
 
 +
</article>
 +
</div>
 +
 
 +
 
 +
 
 +
 
 +
 
 +
 
 +
 
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</div>
 +
 
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<div class="container">
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<div class="row">
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<div class="12u">
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              <a href="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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Revision as of 02:04, 27 October 2017

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Fluorescein Fluorescence standard curve

Plate reader

microplate reader FLUOstar Omega
emission filter: 520 nm
excitation filter: 485 nm

Material

Fluorescein sodium salt
1xPBS
Tissue culture testplate (black with flat bottom)

Method

  1. Prepare fluorescein stock solution

1. Spin down fluorescein stock tube to make sure pellet is at the bottom of tube.
2. Prepare 2x fluorescein stock solution (100 μM) by resuspending fluorescein in 1 mL of 1xPBS.
3. Dilute the 2x fluorescein stock solution with 1xPBS to make a 1x fluorescein solution and resulting concentration of fluorescein stock solution 50 μM

  1. Serial dilutions

1. Add 100 μl of PBS into wells A2, B2, C2, D2....A12, B12, C12, D12
2. Add 200 μl of fluorescein 1x stock solution into A1, B1, C1, D1
3. Transfer 100 μl of fluorescein stock solution from A1 into A2.
4. Mix A2 by pipetting up and down 3x and transfer 100 μl into A3.
5. Mix A3 by pipetting up and down 3x and transfer 100 μl into A4.
6. Mix A4 by pipetting up and down 3x and transfer 100 μl into A5.
7. Mix A5 by pipetting up and down 3x and transfer 100 μl into A6.
8. Mix A6 by pipetting up and down 3x and transfer 100 μl into A7.
9. Mix A7 by pipetting up and down 3x and transfer 100 μl into A8.
10. Mix A8 by pipetting up and down 3x and transfer 100 μl into A9.
11. Mix A9 by pipetting up and down 3x and transfer 100 μl into A10.
12. Mix A10 by pipetting up and down 3x and transfer 100 μl into A11.
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.)

  1. repeat serial dilute for Row B、D、E
  1. Measure fluorescence of all samples in all standard measurement modes in instrument Record the data in your notebook
  1. Import data into Excel (fluorescein standard curve tab ) Sheet_1 provided


Data result







OD600 Reference point

Plate reader

Thermo Scientific™ Multiskan™ FC Filter-based Microplate Photometer
Filter: 595 nm

Material

1 ml LUDOX
mQH2O
96 well cell culture plate (clear with flat-bottom)

Method

1. Add 100 μl LUDOX into wells A1, B1, C1, D1 (or 1 mL LUDOX into cuvette)
2. Add 100 μl of H2O into wells A2, B2, C2, D2 (or 1 mL H2O into cuvette)
3. Measure absorbance 600 nm of all samples in all standard measurement modes in instrument
4. Record the data in excel and Import data into Excel ( OD600 reference point tab ) Sheet_1 provided

Data result



Cell measure

Material

Competent cells ( Escherichia coli strain DH5α)
LB (Luria Bertani) media
Chloramphenicol (stock concentration 25 mg/mL dissolved in EtOH - working stock 25 μg /mL)
50 ml Falcon tube (or equivalent, preferably amber or covered in foil to block light)
Incubator at 37°C
1.5 ml eppendorf tubes for sample storage
Ice bucket with ice
Pipettes
96 well plate(cell culture 96 well plate、tissue culture testplate)
Devices (from InterLab Measurement Kit):
1. Negative control(BBa_R0040)
2. Positive control(J23151+B0032+E0040+B0010+B0012)
3. Test Device 1: J23101+I13504
4. Test Device 2: J23106+I13504
5. Test Device 3: J23117+I13504
6. Test Device 4: J23101+BCD2+E0040+B0015
7. Test Device 5: J23106+BCD2+E0040+B0015
8. Test Device 6: J23117+BCD2+E0040+B0015

Method

1. Day 1 : Resuspended each plasmid in plate 7 and transform into Escherichia coli DH5α.
 (Transformation protocol is from iGEM)
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.
3. Day 3 : Set instrument to read OD600 (as OD calibration setting)and measure OD600 of the overnight cultures
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).
5. Incubate the cultures at 37°C and 170 rpm.
6. Take 1000 μL samples of the cultures at 0, 2, 4, and 6 hours of incubation and place samples on ice
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

Data result