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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: 16px;"> <span style="font-size: 24px"> Western</span> blot identification of proteins<br> CA2L203K CA2WT</p> <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: 16px;"> <span style="font-size: 24px"> Western</span> blot identification of proteins<br> CA2L203K CA2WT</p> <br> | ||
− | <div align="center"><img src="https://static.igem.org/mediawiki/2018/6/60/T--AHUT_China--93.1.png" width="971" height="167" alt=""/></div> | + | <div align="center"><img src="https://static.igem.org/mediawiki/2018/6/60/T--AHUT_China--93.1.png" width="971" height="167" alt=""/></div> <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="754" height="343" alt=""/></div> | + | <div align="center"><img src="https://static.igem.org/mediawiki/2018/8/8b/T--AHUT_China--93.3.png" width="754" height="343" alt=""/></div><br> |
+ | <div class="container"> | ||
+ | <div class="section_title "> | ||
+ | <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;"> <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: 16px;"> <span style="font-size: 24px"> Colorimetric</span> enzyme activity<br> Add 250 μl saturated CO2 solution</p></p> | ||
+ | <br> | ||
+ | <div align="center"><img src="https://static.igem.org/mediawiki/2018/c/c4/T--AHUT_China--95.1.png" width="789" height="328" alt=""/></div> | ||
+ | <div align="center"><img src="https://static.igem.org/mediawiki/2018/8/82/T--AHUT_China--95.2.png" width="669" height="347" alt=""/></div> | ||
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<!--================ Recent Area =================--> | <!--================ Recent Area =================--> |
Revision as of 11:52, 10 October 2018
8.29
Pick Clon
![](https://static.igem.org/mediawiki/2018/0/02/T--AHUT_China--829.png)
8.30
Pilot Expression of Protein
“IPTG+ 4h”“IPTG- 4h”
![](https://static.igem.org/mediawiki/2018/4/44/T--AHUT_China--830.png)
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.
8.31
Protein extraction
CA2L203K
![](https://static.igem.org/mediawiki/2018/2/2d/T--AHUT_China--831.11.png)
![](https://static.igem.org/mediawiki/2018/e/eb/T--AHUT_China--831.12.png)
CA2WT
CA2L203K
![](https://static.igem.org/mediawiki/2018/e/ee/T--AHUT_China--831.2.png)
![](https://static.igem.org/mediawiki/2018/3/3f/T--AHUT_China--831.22.png)
9.01
Purification of Protein
Prior to purification
![](https://static.igem.org/mediawiki/2018/c/cc/T--AHUT_China--91.1.png)
![](https://static.igem.org/mediawiki/2018/7/7b/T--AHUT_China--91.2.png)
Purified
![](https://static.igem.org/mediawiki/2018/8/8a/T--AHUT_China--91.3.png)
![](https://static.igem.org/mediawiki/2018/6/65/T--AHUT_China--91.4.png)
9.03
Western blot identification of proteins
CA2L203K CA2WT
![](https://static.igem.org/mediawiki/2018/6/60/T--AHUT_China--93.1.png)
9.04
Quantification of protein concentrations
![](https://static.igem.org/mediawiki/2018/6/6e/T--AHUT_China--93.2.png)
![](https://static.igem.org/mediawiki/2018/8/8b/T--AHUT_China--93.3.png)
9.05
Colorimetric enzyme activity
Add 250 μl saturated CO2 solution
![](https://static.igem.org/mediawiki/2018/c/c4/T--AHUT_China--95.1.png)
![](https://static.igem.org/mediawiki/2018/8/82/T--AHUT_China--95.2.png)