Difference between revisions of "Team:AHUT China/Notebook"

 
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                                     <li class="nav-item"><a class="nav-link" href="https://2018.igem.org/Team:AHUT_China/INTEGRATED_FOR_GOLD">Integrated HP for gold</a></li>
 
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https://2018.igem.org/Team:AHUT_China/Public_Engagement">Public Engagement</a></li>
 
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                <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-style: normal; font-weight: 400; font-size: 36px; text-align: center;">  &nbsp;&nbsp;&nbsp;<strong style="font-family: Segoe, 'Segoe UI', 'DejaVu Sans', 'Trebuchet MS', Verdana, sans-serif; font-style: normal; font-weight: 400;color: #000000;">Notebook</strong></h2>
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                <hr>
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                <nbsp><nbsp><br><br><br>
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<p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆';  font-size: 14px;"> <span style="font-size: 18px">Wild-type CA2</span> </p></h2><hr>
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<br><br>
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<p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-style: normal; font-weight: 400; font-size: 36px; text-align: center;"> &nbsp;&nbsp;&nbsp;<strong style="font-family: Segoe, 'Segoe UI', 'DejaVu Sans', 'Trebuchet MS', Verdana, sans-serif; font-style: normal; font-weight: 400;color: #000000;"> 8.21</strong></h2>
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The details of the experimental record are as follows:<br></p>
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<iframe src="https://static.igem.org/mediawiki/2018/9/9b/T--AHUT_China--Notebook.pdf" width="1000" height="800"></iframe>
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                <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-style: normal; font-weight: 400; font-size: 36px; text-align: center;">  &nbsp;&nbsp;&nbsp;<strong style="font-family: Segoe, 'Segoe UI', 'DejaVu Sans', 'Trebuchet MS', Verdana, sans-serif; font-style: normal; font-weight: 400;color: #000000;"> 8.29</strong></h2>
 
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                  <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆';  font-size: 14px;"> <span style="font-size: 18px">&nbsp;&nbsp;&nbsp;&nbsp;<span style="font-size: 24px">Pick</span> Clon</span></p>
 
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<br><br>
 
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  <div align="center"><img src="https://static.igem.org/mediawiki/2018/0/02/T--AHUT_China--829.png" width="650"  alt=""/></div><br>
 
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                  <p style="text-align: center; font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 36px;"><strong style="font-family: Segoe, 'Segoe UI', 'DejaVu Sans', 'Trebuchet MS', Verdana, sans-serif; font-style: normal; font-weight: 400; color: #000000;">8.30</strong></p>
 
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                  <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px; ">&nbsp;&nbsp;&nbsp;&nbsp;<span style="font-size: 24px">Pilot</span> Expression of Protein<br>&nbsp;&nbsp;&nbsp;“IPTG+ 4h”“IPTG- 4h”</p> 
 
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<p>&nbsp;</p> <p>&nbsp;</p>             
 
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  <div align="center"><img src="https://static.igem.org/mediawiki/2018/4/44/T--AHUT_China--830.png" width="650"  alt=""/></div><br><br><br><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;">&nbsp;&nbsp;&nbsp;After the rocker culture is 4 H, add IPTG(final concentration is 0.5 mm) and then the rocker culture is 4 hours. Each of the two tubes absorbs 200 μL of bacterial fluid, and in the frozen centrifuge, it is 4 °C and 12000 rpm centrifuge. 10 minutes, go to the clear, It is resuspended with 100 μL 1x SDS Loading buffer, then heated in a water bath pot at 100 °C for 10 minutes and stored in a -20 °C refrigerator.</p>
 
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                <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-style: normal; font-weight: 400; font-size: 36px; text-align: center;">  &nbsp;&nbsp;&nbsp;<strong style="font-family: Segoe, 'Segoe UI', 'DejaVu Sans', 'Trebuchet MS', Verdana, sans-serif; font-style: normal; font-weight: 400;color: #000000;">8.31</strong></h2>
 
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                  <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆';  font-size: 18px;">&nbsp;&nbsp;&nbsp;<span style="font-size: 24px">&nbsp;Protein</span> extraction <br>&nbsp;&nbsp;&nbsp;&nbsp;CA2L203K</p> 
 
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<p>&nbsp;</p> <p>&nbsp;</p> 
 
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  <div align="center"><img src="https://static.igem.org/mediawiki/2018/2/2d/T--AHUT_China--831.11.png" width="800"  alt=""/></div>
 
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<br><br>
 
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<div align="center"><img src="https://static.igem.org/mediawiki/2018/e/eb/T--AHUT_China--831.12.png" width="800"  alt=""/></div>
 
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<p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆';  font-size: 18px;">&nbsp;&nbsp;&nbsp;&nbsp;CA2WT<br>&nbsp;&nbsp;&nbsp;&nbsp;CA2L203K</p>
 
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<br><br>
 
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<div align="center"><img src="https://static.igem.org/mediawiki/2018/e/ee/T--AHUT_China--831.2.png" width="998" alt=""/></div>
 
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<br><br>
 
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<div align="center"><img src="https://static.igem.org/mediawiki/2018/3/3f/T--AHUT_China--831.22.png" width="800"  alt=""/></div>
 
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<br><br><br><br><br>
 
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                <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-style: normal; font-weight: 400; font-size: 36px; text-align: center;">  &nbsp;&nbsp;&nbsp;<strong style="font-family: Segoe, 'Segoe UI', 'DejaVu Sans', 'Trebuchet MS', Verdana, sans-serif; font-style: normal; font-weight: 400;color: #000000;"> 9.01</strong></h2>
 
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                  <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆';  font-size: 14px;"> <span style="font-size: 18px">&nbsp;&nbsp;&nbsp;&nbsp;<span style="font-size: 24px">Purification</span>&nbsp;of Protein <br>&nbsp;&nbsp;&nbsp;&nbsp;Prior to purification</p>
 
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<br>
 
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  <div align="center"><img src="https://static.igem.org/mediawiki/2018/c/cc/T--AHUT_China--91.1.png" width="974"  alt=""/></div>
 
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<br><br>
 
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  <div align="center"><img src="https://static.igem.org/mediawiki/2018/7/7b/T--AHUT_China--91.2.png" width="800"  alt=""/></div>
 
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<br><br><br>
 
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  <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆';  font-size: 14px;"> <span style="font-size: 18px">Purified</p><br><br><br>
 
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<div align="center"><img src="https://static.igem.org/mediawiki/2018/8/8a/T--AHUT_China--91.3.png" width="564" height="75" alt=""/></div>
 
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  <div align="center"><img src="https://static.igem.org/mediawiki/2018/6/65/T--AHUT_China--91.4.png" width="450"  alt=""/></div><br>
 
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                <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-style: normal; font-weight: 400; font-size: 36px; text-align: center;">  &nbsp;&nbsp;&nbsp;<strong style="font-family: Segoe, 'Segoe UI', 'DejaVu Sans', 'Trebuchet MS', Verdana, sans-serif; font-style: normal; font-weight: 400;color: #000000;">9.03</strong></h2>
 
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                  <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆';  font-size: 18px;">&nbsp;&nbsp;&nbsp;<span style="font-size: 24px">&nbsp;Western</span> blot identification of proteins<br>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;CA2L203K&nbsp;&nbsp;&nbsp;&nbsp;CA2WT</p>
 
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<br><br><br>
 
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<div align="center"><img src="https://static.igem.org/mediawiki/2018/6/60/T--AHUT_China--93.1.png" width="9"  alt=""/></div> <br>
 
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<div class="container">
 
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                <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-style: normal; font-weight: 400; font-size: 36px; text-align: center;">  &nbsp;&nbsp;&nbsp;<strong style="font-family: Segoe, 'Segoe UI', 'DejaVu Sans', 'Trebuchet MS', Verdana, sans-serif; font-style: normal; font-weight: 400;color: #000000;">9.04</strong></h2>
 
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                <hr>
 
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                <nbsp><nbsp>
 
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                  <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆';  font-size: 18px;">&nbsp;&nbsp;&nbsp;<span style="font-size: 24px">&nbsp;Quantification</span> of protein concentrations</p> 
 
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<div align="center"><img src="https://static.igem.org/mediawiki/2018/6/6e/T--AHUT_China--93.2.png" width="908"  alt=""/></div>
 
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<br><br>
 
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<div align="center"><img src="https://static.igem.org/mediawiki/2018/8/8b/T--AHUT_China--93.3.png" width="850"  alt=""/></div><br>
 
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<div class="container">
 
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                <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-style: normal; font-weight: 400; font-size: 36px; text-align: center;">  &nbsp;&nbsp;&nbsp;<strong style="font-family: Segoe, 'Segoe UI', 'DejaVu Sans', 'Trebuchet MS', Verdana, sans-serif; font-style: normal; font-weight: 400;color: #000000;">9.05</strong></h2>
 
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                  <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆';  font-size: 18px;">&nbsp;&nbsp;&nbsp;<span style="font-size: 24px">&nbsp;Colorimetric</span> enzyme activity<br>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Add 250 μl saturated CO2 solution</p> 
 
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<br><br>
 
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<div align="center"><img src="https://static.igem.org/mediawiki/2018/c/c4/T--AHUT_China--95.1.png" width="800"  alt=""/></div>
 
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<br><br><br>
 
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<div align="center"><img src="https://static.igem.org/mediawiki/2018/8/82/T--AHUT_China--95.2.png" width="800"  alt=""/></div>
 
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<br><br><br>
 
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<p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆';  font-size: 18px;">&nbsp;&nbsp;&nbsp;According to the average calculated in the table above, the unit enzyme activity of the mutated carbonic anhydride enzyme is 0.3710(10 5 WAU/μg), and the unit enzyme activity of wild carbonic anhydride enzyme is 0.1755(10 5 WAU/μg).<br>
 
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Test esterase activity<br>
 
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Determination curve without carbonated anhydrase:</p> <br>
 
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<br><br>
 
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<div align="center"><img src="https://static.igem.org/mediawiki/2018/f/f6/T--AHUT_China--95.3.png" width="900"  alt=""/></div><br><br>
 
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<p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆';  font-size: 18px;">&nbsp;&nbsp;&nbsp;Determination curve with carbonated anhydrase（CA2WT）</p> <br><br>
 
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<div align="center"><img src="https://static.igem.org/mediawiki/2018/e/e8/T--AHUT_China--95.4.png" width="900"  alt=""/></div>
 
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<p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆';  font-size: 18px;">&nbsp;&nbsp;&nbsp;Determination curve with carbonated anhydrase（CA2L203K）</p> <br><br>
 
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<div align="center"><img src="https://static.igem.org/mediawiki/2018/0/0c/T--AHUT_China--95.5.png" width="900" alt=""/></div>
 
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<br><br><br>
 
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<p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆';  font-size: 18px;">&nbsp;&nbsp;&nbsp;Wild carbonic anhydrase activity calculated as 0.5098 mol/g × min<br>
 
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&nbsp;&nbsp;&nbsp;The mutated carbonic anhydrase enzyme activity was calculated to be 1.0392 mol/g * min.
 
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Latest revision as of 14:40, 17 October 2018

Royal Hotel

Royal Hotel

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      The details of the experimental record are as follows: