Difference between revisions of "Team:HUST-China/InterLab"

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                     <h3 style="background-color:#000733;color:#ffffff; padding:10px;letter-spacing:1px;">Introduction</h3>
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                     <h3 style="background-color:#000733;color:#ffffff; padding:10px;letter-spacing:1px;margin-top:0;">Introduction</h3>
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                     <p>Try to scroll this section and look at the navigation list while scrolling!</p>
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                     <p style="padding:10px">It is significant for synthetic biology to develop a reliable and repeatable measurement, the same to all the other engineering disciplines. We HUST-China have volunteered to test some RBS devices (BCDs) that are intended to make gene expression more precise and reliable by measure the expression level of GFP, in order to help the iGEM community collect data about how reliable will these devices turn out to be in labs around the world.
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                 <div id="section2" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;">  
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                     <h3 style="background-color:#000733;color:#ffffff; padding:10px;letter-spacing:1px;">Provenance and Release</h3>
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                     <h3 style="background-color:#000733;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Provenance and Release</h3>
−
                     <p>Try to scroll this section and look at the navigation list while scrolling!</p>
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                    <h4><strong>①Individuals responsible for conducting InterLab study</strong> </h4>
 +
                     <div class="table-responsive" style="padding: 10px 100px; text-align: center;">
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                        <table class="table">
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                          <thead>
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                              <tr>
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                                <th>Individuals</th>
 +
                                <th>Interlab Part</th>
 +
                              </tr>
 +
                          </thead>
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                          <tbody>
 +
                              <tr>
 +
                                <td>Kangyuan Yu, Haibo Huang, Ziyang Xiao, Shaofeng Liao</td>
 +
                                <td>created the devices</td>
 +
                              </tr>
 +
                              <tr>
 +
                                <td>Efan Wang, Long Cheng, HuiPing Shi </td>
 +
                                <td>conducted the measurements</td>
 +
                              </tr>
 +
                              <tr>
 +
                                <td>Efan Wang</td>
 +
                                <td>processed the data</td>
 +
                              </tr>
 +
                          </tbody>
 +
                        </table>
 +
                    </div>
 +
                    <h4><strong> ②Corresponding email</strong> </h4>
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                    <div class="table-responsive" style="padding: 10px 100px; text-align: center;">
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                        <table class="table">
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                          <thead>
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                              <tr>
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                                <th>Efan Wang </th>
 +
                                <th>erfan@hust.edu.cn</th>
 +
                              </tr>
 +
                          </thead>
 +
                          <tbody>
 +
                              <tr>
 +
                                <td>Kangyuan Yu</td>
 +
                                <td>985930862@qq.com</td>
 +
                              </tr>
 +
                              <tr>
 +
                                <td>Haibo Huang </td>
 +
                                <td>u201512127@hust.edu.cn</td>
 +
                              </tr>
 +
                              <tr>
 +
                                <td>Ziyang Xiao</td>
 +
                                <td>372657289@qq.com</td>
 +
                              </tr>
 +
                              <tr>
 +
                                <td>Shaofeng Liao</td>
 +
                                <td> 15827233830@qq.com</td>
 +
                              </tr>
 +
                              <tr>
 +
                                <td>HuiPing Shi</td>
 +
                                <td>172295915@qq.com</td>
 +
                              </tr>
 +
                              <tr>
 +
                                <td>Long Cheng</td>
 +
                                <td>u201512127@hust.edu.cn</td>
 +
                              </tr>
 +
                          </tbody>
 +
                        </table>
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                    </div>
 
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                     <h3 style="background-color:#000733;color:#ffffff; padding:10px;letter-spacing:1px;">Chassis and Safety Information</h3>
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                     <h3 style="background-color:#000733;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Chassis and Safety Information</h3>
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                     <p>Try to scroll this section and look at the navigation list while scrolling!</p>
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                    <h4><strong> What chassis did you use?</strong> </h4>
 +
                     <p>Escherichia coli DH5alpha</p>
 +
                    <h4><strong> What Biosafety Level is your chassis? </strong> </h4>
 +
                    <p>BSL1</p>
 +
                    <h4><strong>What PPE did you utilize during your experiments? </strong></h4>
 +
                    <p>Tianming gloves</p>
 +
                    <p>Songxinjiujiu labcoats</p>
 
                 </div>
 
                 </div>
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                     <h3 style="background-color:#000733;color:#ffffff; padding:10px;letter-spacing:1px;">Instrument Information</h3>
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                     <h3 style="background-color:#000733;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Instrument Information</h3>
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                     <p>Try to scroll this section and look at the navigation list while scrolling!</p>
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                    <h4><strong> hat instrument did you use during your measurements? </strong> </h4>
 +
                     <p>plate reader</p>
 +
                    <h4><strong> Please provide the brand and model of your instrument.</strong> </h4>
 +
                    <p>Flexstation 3</p>
 
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                     <h3 style="background-color:#000733;color:#ffffff; padding:10px;letter-spacing:1px;">Calibration Protocol</h3>
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                     <h3 style="background-color:#000733;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Calibration Protocol</h3>
−
                     <p>Try to scroll this section and look at the navigation list while scrolling!</p>
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 +
 
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                    <h4><strong>  A1. Protocol for Optical Density (OD600) Standard Measurement</strong> </h4>
 +
                    <h5><strong>Did you use pathlength correction during measurement? </strong> </h5>
 +
                     <p>Yes</p>
 +
                    <h5><strong>Number of flashes per well  </strong> </h5>
 +
                    <p>6</p>
 +
                    <h5><strong>Orbital averaging (mm)  </strong> </h5>
 +
                    <p>600</p>
 +
                    <h5><strong> What temperature setting did you use during the measurement? </strong> </h5>
 +
                    <p>22℃</p>
 +
                    <h5><strong>What type of 96-well plate did you use? </strong> </h5>
 +
                    <p>Black plate (preferred)</p>
 +
                    <h5><strong> Did your plate have flat-bottomed or round-bottomed wells? </strong> </h5>
 +
                    <p>Flat</p>
 +
 
 +
 
 +
                    <h4><strong> A2. Measurement Steps</strong>
 +
                        <li class="dropdown" style="list-style-type:none;">
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                            <ul class="dropdown-menu" style="padding: 10px;">
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                                <li>① Prepare your 96 well plate</li>
 +
                                <li>②Add 100 μl LUDOX 100 % into wells A1, B1, C1, D1</li>
 +
                                <li>③Add 100 μl of dH2O into A2, B2, C2, D2</li>
 +
                                <li>④Measure absorbance 600 nm of all samples in all standard measurement modes in instrument</li>
 +
                                <li>⑤Import data into "Abs600" blue cells in provided Excel calibration sheet</li>
 +
                            </ul>
 +
                        </li>
 +
                    </h4>
 +
                   
 +
                    <h4><strong>  B1. Protocol for FIuorescein Fluoresence standard curve</strong> </h4>
 +
                    <h5><strong>Did you use pathlength correction during measurement? </strong> </h5>
 +
                    <p>Yes</p>
 +
                    <h5><strong>Number of flashes per well  </strong> </h5>
 +
                    <p>6</p>
 +
                    <h5><strong>What gain setting did you use?  </strong> </h5>
 +
                    <p>Automatic</p>
 +
                    <h5><strong> If you used a filter, what light wavelengths did it pass?  </strong> </h5>
 +
                    <p>530nm</p>
 +
                    <h5><strong>Emission wavelength </strong> </h5>
 +
                    <p>530nm</p>
 +
                    <h5><strong> Excitation wavelength </strong> </h5>
 +
                    <p>485nm</p>
 +
                    <h5><strong> Fluorescence reading  </strong> </h5>
 +
                    <p>Bottom optic</p>
 +
                    <h5><strong>What type of 96-well plate did you use? </strong> </h5>
 +
                    <p>Black plate (preferred)</p>
 +
                    <h5><strong> Did your plate have flat-bottomed or round-bottomed wells? </strong> </h5>
 +
                    <p>Flat</p>
 +
                    <h5><strong> What temperature setting did you use during the measurement?  </strong> </h5>
 +
                    <p>22℃</p>
 +
 
 +
                    <h4><strong> A2. Measurement Steps</strong></h4>
 +
                    <h5><strong>  Part 1: Prepare the Fluorescein stock solution </strong> </h5>
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                            <ul class="dropdown-menu" style="padding: 10px;">
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                                <li>①  Spin down fluorescein stock tube to make sure pellet is at the bottom of tube.</li>
 +
                                <li>②  Prepare 2x fluorescein stock solution (100 μM) by resuspending fluorescein in 1ml of 1xPBS</li>
 +
                                <li>③  Dilute the 2x fluorescein stock solution using 1xPBS to make a 1x fluorescein solution (final concentration is 50 μM).</li>
 +
                                <li>④  *Illustration of serial dilution samples in 96 well plate or cuvettes: value decreases by 2-fold with each column (50% in column 2, 25% in column 3, 12.5% in column 4, etc.)</li>
 +
                                <li>⑤Import data into "Abs600" blue cells in provided Excel calibration sheet</li>
 +
                            </ul>
 +
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 +
 
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Revision as of 14:35, 14 October 2017

Interlab

「Interlab」

Introduction

It is significant for synthetic biology to develop a reliable and repeatable measurement, the same to all the other engineering disciplines. We HUST-China have volunteered to test some RBS devices (BCDs) that are intended to make gene expression more precise and reliable by measure the expression level of GFP, in order to help the iGEM community collect data about how reliable will these devices turn out to be in labs around the world.

Provenance and Release

①Individuals responsible for conducting InterLab study

Individuals Interlab Part
Kangyuan Yu, Haibo Huang, Ziyang Xiao, Shaofeng Liao created the devices
Efan Wang, Long Cheng, HuiPing Shi conducted the measurements
Efan Wang processed the data

②Corresponding email

Efan Wang erfan@hust.edu.cn
Kangyuan Yu 985930862@qq.com
Haibo Huang u201512127@hust.edu.cn
Ziyang Xiao 372657289@qq.com
Shaofeng Liao 15827233830@qq.com
HuiPing Shi 172295915@qq.com
Long Cheng u201512127@hust.edu.cn

Chassis and Safety Information

What chassis did you use?

Escherichia coli DH5alpha

What Biosafety Level is your chassis?

BSL1

What PPE did you utilize during your experiments?

Tianming gloves

Songxinjiujiu labcoats

Instrument Information

hat instrument did you use during your measurements?

plate reader

Please provide the brand and model of your instrument.

Flexstation 3

Calibration Protocol

A1. Protocol for Optical Density (OD600) Standard Measurement

Did you use pathlength correction during measurement?

Yes

Number of flashes per well

6

Orbital averaging (mm)

600

What temperature setting did you use during the measurement?

22℃

What type of 96-well plate did you use?

Black plate (preferred)

Did your plate have flat-bottomed or round-bottomed wells?

Flat

A2. Measurement Steps
    • ① Prepare your 96 well plate
    • ②Add 100 μl LUDOX 100 % into wells A1, B1, C1, D1
    • ③Add 100 μl of dH2O into A2, B2, C2, D2
    • ④Measure absorbance 600 nm of all samples in all standard measurement modes in instrument
    • ⑤Import data into "Abs600" blue cells in provided Excel calibration sheet

    • B1. Protocol for FIuorescein Fluoresence standard curve

    Did you use pathlength correction during measurement?

    Yes

    Number of flashes per well

    6

    What gain setting did you use?

    Automatic

    If you used a filter, what light wavelengths did it pass?

    530nm

    Emission wavelength

    530nm

    Excitation wavelength

    485nm

    Fluorescence reading

    Bottom optic

    What type of 96-well plate did you use?

    Black plate (preferred)

    Did your plate have flat-bottomed or round-bottomed wells?

    Flat

    What temperature setting did you use during the measurement?

    22℃

    A2. Measurement Steps

    Part 1: Prepare the 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 1ml of 1xPBS
    • ③ Dilute the 2x fluorescein stock solution using 1xPBS to make a 1x fluorescein solution (final concentration is 50 μM).
    • ④ *Illustration of serial dilution samples in 96 well plate or cuvettes: value decreases by 2-fold with each column (50% in column 2, 25% in column 3, 12.5% in column 4, etc.)
    • ⑤Import data into "Abs600" blue cells in provided Excel calibration sheet

    • Referrence:

    1.Binnemans K, Jones P T, Blanpain B, et al. Recycling of rare earths: a critical review[J]. Journal of Cleaner Production, 2013, 51(14):1–22.

    2.Bradsher, Keith (October 29, 2010). "After China's Rare Earth Embargo, a New Calculus". The New York Times. Retrieved October 30, 2010.