(11 intermediate revisions by 3 users not shown) | |||
Line 309: | Line 309: | ||
} | } | ||
− | @media screen and (max-width: | + | @media screen and (max-width: 992px) { |
#section1, #section2, #section3, #section4, #section5 { | #section1, #section2, #section3, #section4, #section5 { | ||
margin-left: 50px; background-color: #eee; color:#171717; | margin-left: 50px; background-color: #eee; color:#171717; | ||
Line 317: | Line 317: | ||
body { | body { | ||
− | + | position: relative; | |
} | } | ||
+ | @media (min-width: 768px) { | ||
.menu_logo_1 { | .menu_logo_1 { | ||
− | + | top: 18px; | |
− | + | left:10px; | |
− | + | z-index: 1000; | |
− | + | position: fixed; | |
− | + | ||
} | } | ||
.menu_logo_2 { | .menu_logo_2 { | ||
− | + | top: 18px; | |
− | + | left:100px; | |
− | + | z-index: 1000; | |
− | + | position: fixed; | |
− | + | } | |
+ | .nav-pills{ | ||
+ | font-size:8px; | ||
+ | font-weight: bold; | ||
+ | } | ||
+ | } | ||
+ | |||
+ | /* 中等屏幕(桌面显示器,大于等于992px) */ | ||
+ | @media (min-width: 992px) { | ||
+ | .menu_logo_1 { | ||
+ | top: 18px; | ||
+ | left:20px; | ||
+ | z-index: 1000; | ||
+ | position: fixed; | ||
+ | } | ||
+ | .menu_logo_2 { | ||
+ | top: 18px; | ||
+ | left:110px; | ||
+ | z-index: 1000; | ||
+ | position: fixed; | ||
+ | } | ||
+ | .nav-pills{ | ||
+ | font-size:10px; | ||
+ | font-weight: bold; | ||
+ | } | ||
+ | } | ||
+ | |||
+ | /* 大屏幕(大桌面显示器,大于等于1200px) */ | ||
+ | @media (min-width: 1200px) { | ||
+ | .menu_logo_1 { | ||
+ | top: 18px; | ||
+ | left:80px; | ||
+ | z-index: 1000; | ||
+ | position: fixed; | ||
+ | } | ||
+ | .menu_logo_2 { | ||
+ | top: 18px; | ||
+ | left:170px; | ||
+ | z-index: 1000; | ||
+ | position: fixed; | ||
+ | } | ||
+ | .nav-pills{ | ||
+ | font-size:12px; | ||
+ | font-weight: bold; | ||
+ | } | ||
} | } | ||
Line 468: | Line 512: | ||
<li><a href="#section5">Calibration Protocol</a></li> | <li><a href="#section5">Calibration Protocol</a></li> | ||
<li><a href="#section6">Cell Culture</a></li> | <li><a href="#section6">Cell Culture</a></li> | ||
− | <li><a href="#section7">Interlab | + | <li><a href="#section7">Interlab Results</a></li> |
</ul> | </ul> | ||
</div> | </div> | ||
Line 476: | Line 520: | ||
<div id="section1" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;"> | <div id="section1" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;"> | ||
<h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px;margin-top:0;">Introduction</h3> | <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px;margin-top:0;">Introduction</h3> | ||
− | <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 | + | <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. |
</p> | </p> | ||
</div> | </div> | ||
Line 522: | Line 566: | ||
<tr> | <tr> | ||
<td>Kangyuan Yu</td> | <td>Kangyuan Yu</td> | ||
− | <td> | + | <td>u201612160@hust.edu.cn</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Haibo Huang </td> | <td>Haibo Huang </td> | ||
− | <td> | + | <td>u201612177@hust.edu.cn</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Ziyang Xiao</td> | <td>Ziyang Xiao</td> | ||
− | <td> | + | <td>u201612166@hust.edu.cn</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Shaofeng Liao</td> | <td>Shaofeng Liao</td> | ||
− | <td> | + | <td>u201612159@hust.edu.cn</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>HuiPing Shi</td> | <td>HuiPing Shi</td> | ||
− | <td> | + | <td>u201513154@hust.edu.cn</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 551: | Line 595: | ||
<h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Chassis and Safety</h3> | <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Chassis and Safety</h3> | ||
<h4><strong> What chassis did you use?</strong> </h4> | <h4><strong> What chassis did you use?</strong> </h4> | ||
− | <p>Escherichia coli | + | <p>Escherichia coli DH5α</p> |
<h4><strong> What Biosafety Level is your chassis? </strong> </h4> | <h4><strong> What Biosafety Level is your chassis? </strong> </h4> | ||
<p>BSL1</p> | <p>BSL1</p> | ||
Line 561: | Line 605: | ||
<h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Instrument</h3> | <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Instrument</h3> | ||
<h4><strong> What instrument did you use during your measurements? </strong> </h4> | <h4><strong> What instrument did you use during your measurements? </strong> </h4> | ||
− | <p> | + | <p>plate reader</p> |
<h4><strong> Please provide the brand and model of your instrument.</strong> </h4> | <h4><strong> Please provide the brand and model of your instrument.</strong> </h4> | ||
<p>Flexstation 3</p> | <p>Flexstation 3</p> | ||
Line 593: | Line 637: | ||
<li>②Add 100 μl LUDOX 100 % into wells A1, B1, C1, D1</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>③Add 100 μl of dH2O into A2, B2, C2, D2</li> | ||
− | <li>④Measure absorbance 600 nm of all samples in all standard measurement | + | <li>④Measure absorbance 600 nm of all samples in all standard measurement modes in instrument</li> |
− | + | ||
</ul> | </ul> | ||
</li> | </li> | ||
− | <h4><strong> B1. Protocol for | + | <h4><strong> B1. Protocol for FIuorescein Fluoresence standard curve</strong> </h4> |
<h5><strong>Did you use pathlength correction during measurement? </strong> </h5> | <h5><strong>Did you use pathlength correction during measurement? </strong> </h5> | ||
<p>Yes</p> | <p>Yes</p> | ||
Line 632: | Line 675: | ||
<li>③ Dilute the 2x fluorescein stock solution using 1xPBS to make a 1x fluorescein solution (final concentration is 50 μM).</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>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> | ||
− | |||
</ul> | </ul> | ||
</li> | </li> | ||
Line 642: | Line 684: | ||
<div class="col-xs-12" > | <div class="col-xs-12" > | ||
<h5><strong> Part 2: Prepare the serial dilutions of Fluorescein </strong> </h5> | <h5><strong> Part 2: Prepare the serial dilutions of Fluorescein </strong> </h5> | ||
− | < | + | <ul style="list-style: none; font-size:10px;"> |
<li>①Add 100 μl of 1xPBS into wells A2, B2, C2, D2....A12, B12, C12, D12.</li> | <li>①Add 100 μl of 1xPBS into wells A2, B2, C2, D2....A12, B12, C12, D12.</li> | ||
<li>②Add 200 μl of 1x Fluorescein stock solution into A1, B1, C1, D1.</li> | <li>②Add 200 μl of 1x Fluorescein stock solution into A1, B1, C1, D1.</li> | ||
Line 659: | Line 701: | ||
<li>⑮ Repeat dilution series for rows B, C, D. | <li>⑮ Repeat dilution series for rows B, C, D. | ||
<li>⑯ Measure fluorescence of all samples in all standard measurement modes in instrument. | <li>⑯ Measure fluorescence of all samples in all standard measurement modes in instrument. | ||
− | </ | + | </ul> |
</div> | </div> | ||
Line 706: | Line 748: | ||
<li>① Set your instrument to read Abs600 (as OD calibration setting)</li> | <li>① Set your instrument to read Abs600 (as OD calibration setting)</li> | ||
<li>② Measure Abs600 of the overnight cultures</li> | <li>② Measure Abs600 of the overnight cultures</li> | ||
− | <li>③ | + | <li>③ Dilute the cultures to a target Abs600 of 0.02 (see the volume of preloading culture and media in Excel Dilution Calculation sheets) in 12 ml LB medium + Chloramphenicol in 50 mL falcon tube.</li> |
− | + | <li>④ Remove 500uL of each culture for your zero time point and hold these samples on ice. (You should have 16 sample tubes from the time=0 time point)</li> | |
− | <li> | + | <li>⑤ Incubate the cultures at 37°C and 220 rpm.</li> |
− | <li> | + | <li>⑥ Remove 500uL samples of each culture after 2, 4 and 6 hours of growth. Keep samples on ice while completing the additional time points. You should have 16 sample tubes for each time point.</li> |
− | <li> | + | <li>⑦ Set up your four measurement plates: For colony #1 culture, pipet 100 uL samples into wells A, B, C and D of the row for that device. For colony #2, pipet 100uL samples into wells E, F, G, and H of the row for that device.</li> |
− | <li> | + | <li>⑧ Read your plates, taking care to use the exact same settings used for your fluorescein measurement.</li> |
− | <li> | + | |
Line 734: | Line 775: | ||
<ul class="yuandian"> | <ul class="yuandian"> | ||
<li>Measure OD and fluorescence of all samples</li> | <li>Measure OD and fluorescence of all samples</li> | ||
− | |||
</ul> | </ul> | ||
<h5><strong>Suggested Plate Layout for 96-well Plate</strong> </h5> | <h5><strong>Suggested Plate Layout for 96-well Plate</strong> </h5> | ||
Line 744: | Line 784: | ||
</div> | </div> | ||
<div id="section7" style="border: solid 1px #666; margin:5px; padding:5px;border-radius:10px;overflow: hidden;"> | <div id="section7" style="border: solid 1px #666; margin:5px; padding:5px;border-radius:10px;overflow: hidden;"> | ||
− | <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px;margin-top:0;">Interlab | + | <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px;margin-top:0;">Interlab Resultn</h3> |
<h5 class="col-xs-12"><strong>OD600 Reference Point</strong> </h5> | <h5 class="col-xs-12"><strong>OD600 Reference Point</strong> </h5> | ||
<img title="demo1" src="https://static.igem.org/mediawiki/2017/5/52/2017_HUST_China_interlab_img_table2.png" alt="demo1" class="col-xs-8 col-xs-offset-2"> | <img title="demo1" src="https://static.igem.org/mediawiki/2017/5/52/2017_HUST_China_interlab_img_table2.png" alt="demo1" class="col-xs-8 col-xs-offset-2"> |
Latest revision as of 18:00, 1 November 2017