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| {{:Team:Tianjin/Templates/MaterialTheme|}} | | {{:Team:Tianjin/Templates/MaterialTheme|}} |
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− | {{:Team:Tianjin/Templates/AddCSS|:Team:Tianjin/Note/CFPS/camart.css}} | + | {{:Team:Tianjin/Templates/AddCSS|:Team:Tianjin/Note/R-R/camarts.css}} |
| {{:Team:Tianjin/Templates/AddCSS|:Team:Tianjin/Experiment/css/font.css}} | | {{:Team:Tianjin/Templates/AddCSS|:Team:Tianjin/Experiment/css/font.css}} |
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| <html lang="en"> | | <html lang="en"> |
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| + | .note-content,.note-content2,.note-content3,.note-content4,.note-content5,.note-content6,.note-content li,.note-content2 li,.note-content3 li,.note-content4 li,.note-content5 li,.note-content6 li{ |
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| <div id="main"> | | <div id="main"> |
| <div id="content" class="home blog single-author one-column content" role="main"> | | <div id="content" class="home blog single-author one-column content" role="main"> |
− | <article id="post-4252" class="post-4252 post type-post status-publish format-standard has-post-thumbnail hentry category-150 tag-174 tag-xinjiang tag-173">
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| + | <!--------------------------------week1 start--------------------------------------------> |
| + | <div id="Week1"></div> |
| + | <article id="post-4252" class="post-4252 post type-post status-publish format-standard has-post-thumbnail hentry category-150 tag-174 tag-xinjiang tag-173"> |
| <header class="entry-header"> | | <header class="entry-header"> |
− | <div class="entry-title" align="center" ><a href="https://www.camarts.cn/archives/4252.html" title="查看《火影忍者疾风传》中的全部作品" rel="bookmark">Notes</a></div> | + | <div class="entry-title" align="center" >Notes For CFPS system</div> |
| </header><!-- .entry-header --> | | </header><!-- .entry-header --> |
− | <h1 class="entry-title">Week1(8/1/2016-8/7/2016)</h1> | + | <h1 class="entry-title">Week1(8/24/2016-8/30/2016)</h1> |
| <div class="entry-content"> | | <div class="entry-content"> |
− | <b>Ⅰ.Constraction of expression plasmid in CFPS(cell-free protein synthesis)</b><br/> | + | <div class="note-content"> |
− | The mutations of PETase gene had been contracted by over-lap PCR and Restriction Enzyme cutting sites(SacⅠ&EcoRⅠ) had been added to it’s ends (5’&3’) separately. | + | |
| + | <b>Ⅰ.Constraction of expression plasmid in CFPS(cell-free protein synthesis)</b><br/> |
| + | The mutants of PETase gene had been contracted by over-lap PCR and Restriction Enzyme cutting sites(SacⅠ&EcoRⅠ) had been added to it’s ends (5’&3’) separately. |
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− | <hr class="article">
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− | </article><!-- #post-4252 --> | + | </article> #post-4252 --> |
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| + | <!------------------------------------week1 end------------------------------------------------> |
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| + | <!------------------------------------week2 start------------------------------------------------> |
| + | <div id="Week2"></div> |
| <article id="post-4252" class="post-4252 post type-post status-publish format-standard has-post-thumbnail hentry category-150 tag-174 tag-xinjiang tag-173"> | | <article id="post-4252" class="post-4252 post type-post status-publish format-standard has-post-thumbnail hentry category-150 tag-174 tag-xinjiang tag-173"> |
| <header class="entry-header"> | | <header class="entry-header"> |
− | <h1 class="entry-title"><a href="https://www.camarts.cn/archives/4252.html" title="查看《火影忍者疾风传》中的全部作品" rel="bookmark">Week2(8/15/2016-8/21/2016)</a></h1> | + | <h1 class="entry-title">Week2(8/31/2016-9/6/2016)</h1> |
| </header><!-- .entry-header --> | | </header><!-- .entry-header --> |
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| <div class="entry-content"> | | <div class="entry-content"> |
− | <p><li> XbaⅠ&SacⅠdouble restriction endonuclease digestion in <i>CFP</i> gene.</li><br /> | + | |
| + | <div class="note-content2"> |
| + | <p><li> XbaⅠ&SacⅠdouble restriction endonuclease digestion in <i>CFP</i> gene.</li><br /> |
| <li>XbaⅠ&EcoRⅠdouble restriction endonuclease digestion in pRset_CFP-1<br/> | | <li>XbaⅠ&EcoRⅠdouble restriction endonuclease digestion in pRset_CFP-1<br/> |
| Bands as expected(760bp&3000bp) in agarose gel , gel purification of the digested products(3000bp). | | Bands as expected(760bp&3000bp) in agarose gel , gel purification of the digested products(3000bp). |
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| Verification : Colony PCR with fast taq Polymerase, primers: PETase-S/PETase -A, template: Colony of E.coli with plasmid pRset_CFP-1-M154L. Bands as expected(about 760bp) in agarose gel. | | Verification : Colony PCR with fast taq Polymerase, primers: PETase-S/PETase -A, template: Colony of E.coli with plasmid pRset_CFP-1-M154L. Bands as expected(about 760bp) in agarose gel. |
| </li><br /> | | </li><br /> |
− | <li>Cultivation the E.coli cells transformed yesterday in several 5mL liquid LB culture mediums with Amp at 37℃ for 12 hours.<br/>
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− | <li>Plasmid isolation of pRset_CFP-1-M154L.<br/>
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| <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/5/55/T--Tianjin--cell-free_note-1.png" alt="T--Tianjin--cell-free_note-1" width="800" height="533"> | | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/5/55/T--Tianjin--cell-free_note-1.png" alt="T--Tianjin--cell-free_note-1" width="800" height="533"> |
| + | |
| + | <li>Cultivation the E.coli cells transformed yesterday in several 5mL liquid LB culture mediums with Amp at 37℃ for 12 hours.</li><br/> |
| + | <li>Plasmid isolation of pRset_CFP-1-M154L.</li><br/> |
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| </p> | | </p> |
− | <p> <a href="https://www.camarts.cn/archives/4252.html" class="more-link">查看Team Tianjin全部实验 <span class="meta-nav">›</span></a></p> | + | </div> |
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| </div><!-- .entry-content --> | | </div><!-- .entry-content --> |
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− | <footer class="entry-meta">
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− | <span class="sep">发表于 </span>2016 年 1 月 28 日<span class="sep"> | </span> | + | |
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− | <span class="cat-links">
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− | <span class="entry-utility-prep entry-utility-prep-cat-links">发表在</span> <a href="https://www.camarts.cn/archives/category/%e8%a5%bf%e5%8c%97%e8%a1%8c" rel="category tag">春野樱</a> </span>
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− | <span class="sep"> | </span>
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− | <span class="tag-links">
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− | <span class="entry-utility-prep entry-utility-prep-tag-links">标签有</span> <a href="https://www.camarts.cn/archives/tag/%e5%b8%83%e5%b0%94%e6%b4%a5" rel="tag">春野樱</a>、<a href="https://www.camarts.cn/archives/tag/xinjiang" rel="tag">新疆</a>、<a href="https://www.camarts.cn/archives/tag/%e9%98%bf%e5%8b%92%e6%b3%b0" rel="tag">漩涡鸣人</a> </span>
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− | <span class="sep"> | </span>
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− | <span class="total-pictures"><a href="https://www.camarts.cn/archives/4252.html">5 张图片</a></span>
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− | </footer><!-- #entry-meta -->
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− | <hr class="article">
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− | </article>
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| + | <!------------------------------------week2 end------------------------------------------------> |
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− | <article id="post-4252" class="post-4252 post type-post status-publish format-standard has-post-thumbnail hentry category-150 tag-174 tag-xinjiang tag-173">
| + | <!------------------------------------week3 start------------------------------------------------> |
− | <h1 class="entry-title"><a href="https://www.camarts.cn/archives/4252.html" title="查看《火影忍者疾风传》中的全部作品" rel="bookmark">Week3(8/29/2016-9/4/2016)</a></h1> | + | <div id="Week3"></div> |
| + | <article id="post-4252" class="post-4252 post type-post status-publish format-standard has-post-thumbnail hentry category-150 tag-174 tag-xinjiang tag-173"> |
| + | <h1 class="entry-title">Week3(9/7/2016-9/13/2016)</h1> |
| <div class="entry-content"> | | <div class="entry-content"> |
− | <p><li>Our strategy is to exchange the site-directed mutation PETase-M154L for the other 21 mutations and 1 wild type, separately.<br/> | + | |
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| + | <div class="note-content3"> |
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| + | <p><li>Our strategy is to exchange the site-directed mutant PETase-M154L for the other 21 mutants and 1 wild type, separately.<br/> |
| XbaⅠ&EcoRⅠdouble restriction endonuclease digestion in pRset_CFP-1-M154L<br/> | | XbaⅠ&EcoRⅠdouble restriction endonuclease digestion in pRset_CFP-1-M154L<br/> |
| No expected bands(3600bp&750bp) in agarose gel. We don’t find the reason why the plasmid constuced can’t be cut into the two expected bands, so we decided to change the initial strategy. | | No expected bands(3600bp&750bp) in agarose gel. We don’t find the reason why the plasmid constuced can’t be cut into the two expected bands, so we decided to change the initial strategy. |
| </li><br/> | | </li><br/> |
− | <li>Ligaion of digested pRset_CFP-1, digested <i>CFP<i/> gene and the rest 22 digested gene interested (including 21 PETase site-directed mutations geneand 1 wild type ) separately in accordance with the 1:5:5 molecular ratio. The newly constructed plasmid is called pRset_CFP-1-R90I (R90T, R90A, S92A, Q119A, M154A, W159H, W159M, N205L, N205V, S207T, S207A, I208V, S238F, S238V,A240P, S242I.)</li><br/> | + | <li>Ligaion of digested pRset_CFP-1, digested <i>CFP</i> gene and the rest 22 digested gene interested (including 21 PETase site-directed mutants geneand 1 wild type ) separately in accordance with the 1:5:5 molecular ratio. The newly constructed plasmid is called pRset_CFP-1-R90I (R90T, R90A, S92A, Q119A, M154A, W159H, W159M, N205L, N205V, S207T, S207A, I208V, S238F, S238V,A240P, S242I.)</li><br/> |
| <li>Transformation</li> | | <li>Transformation</li> |
| <li> Plasmid isolation</li> | | <li> Plasmid isolation</li> |
| <li> Verification</li> | | <li> Verification</li> |
− | </p> | + | </p> |
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| + | </div> |
| + | <a class="expand-btn3">Show More</a> |
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− | <p> <a href="https://www.camarts.cn/archives/4252.html" class="more-link">查看Team Tianjin全部实验 <span class="meta-nav">›</span></a></p>
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− | <footer class="entry-meta">
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− | <span class="sep">发表于 </span>2016 年 1 月 28 日<span class="sep"> | </span>
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− |
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− | <span class="cat-links">
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− | <span class="entry-utility-prep entry-utility-prep-cat-links">发表在</span> <a href="https://www.camarts.cn/archives/category/%e8%a5%bf%e5%8c%97%e8%a1%8c" rel="category tag">春野樱</a> </span>
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− | <span class="sep"> | </span>
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− | <span class="tag-links">
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− | <span class="entry-utility-prep entry-utility-prep-tag-links">标签有</span> <a href="https://www.camarts.cn/archives/tag/%e5%b8%83%e5%b0%94%e6%b4%a5" rel="tag">春野樱</a>、<a href="https://www.camarts.cn/archives/tag/xinjiang" rel="tag">新疆</a>、<a href="https://www.camarts.cn/archives/tag/%e9%98%bf%e5%8b%92%e6%b3%b0" rel="tag">漩涡鸣人</a> </span>
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− | <span class="sep"> | </span>
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− | <span class="total-pictures"><a href="https://www.camarts.cn/archives/4252.html">5 张图片</a></span>
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| </div><!-- .entry-content --> | | </div><!-- .entry-content --> |
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− | </footer><!-- #entry-meta -->
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− | <hr class="article"> | + | |
− | </article>
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| + | <!------------------------------------week3 end------------------------------------------------> |
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| + | <!------------------------------------week4 start------------------------------------------------> |
| + | <div id="Week4"></div> |
| <article id="post-4252" class="post-4252 post type-post status-publish format-standard has-post-thumbnail hentry category-150 tag-174 tag-xinjiang tag-173"> | | <article id="post-4252" class="post-4252 post type-post status-publish format-standard has-post-thumbnail hentry category-150 tag-174 tag-xinjiang tag-173"> |
− | <h1 class="entry-title"><a href="https://www.camarts.cn/archives/4252.html" title="查看《火影忍者疾风传》中的全部作品" rel="bookmark">Week4(9/5/2016-9/11/2016)</a></h1> | + | <h1 class="entry-title">Week4(9/14/2016-9/20/2016)</h1> |
| <div class="entry-content"> | | <div class="entry-content"> |
− | <b>Ⅱ. Expression of the plasmids constracted before<b> | + | |
− | <p>
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| + | <div class="note-content4"> |
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| + | <b>Ⅱ. Expression of the plasmids constracted before</b> |
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| <li>We choosed 4 tubes of plasmid to express in the cell-free system firstly.<br/> | | <li>We choosed 4 tubes of plasmid to express in the cell-free system firstly.<br/> |
| The protocol of the cell-free protein synthesis system(50 µL) we used :<br/> | | The protocol of the cell-free protein synthesis system(50 µL) we used :<br/> |
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| Gene( plasmid as template) 1µL<br/> | | Gene( plasmid as template) 1µL<br/> |
| Lysate 15µL<br/> | | Lysate 15µL<br/> |
− | (PS: the details of the system are not available)<br/> | + | (PS: The details of the system are not available.)<br/> |
| </li> | | </li> |
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− | <b>Systems were expressed in 96-well plates and put the 96-well plates into the ELIASA (microplate reader) at 30℃ for 12 hours.<b><br/> | + | <b>Systems were expressed in 96-well plates and put the 96-well plates into the ELIASA (microplate reader) at 30℃ for 12 hours.</b><br/> |
| <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/3/3d/T--Tianjin--cell-free_note-2.png" alt="T--Tianjin--cell-free_note-2" width="800" height="533"> | | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/3/3d/T--Tianjin--cell-free_note-2.png" alt="T--Tianjin--cell-free_note-2" width="800" height="533"> |
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− | <li><b>Parallel experiments for expression in CFPS system<b></li><br/> | + | <li><b>Parallel experiments for expression in CFPS system</b></li><br/> |
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− | <div class="note-content">
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− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/9/97/T--Tianjin--cf-blank.png" alt="T--Tianjin--cf-blank" width="800" height="533"> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/9/97/T--Tianjin--cf-blank.png" alt="T--Tianjin--cf-blank" height="250px"> |
− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/d/d0/T--Tianjin--cf-m1.png" alt="T--Tianjin--cf-m1" width="800" height="533"> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/d/d0/T--Tianjin--cf-m1.png" alt="T--Tianjin--cf-m1" height="250px"> |
− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/4/44/T--Tianjin--cf-m2.png" alt="T--Tianjin--cf-m2" width="800" height="533"> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/4/44/T--Tianjin--cf-m2.png" alt="T--Tianjin--cf-m2" height="250px"> |
− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/3/34/T--Tianjin--cf-m3.png" alt="T--Tianjin--cf-m3" width="800" height="533"> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/3/34/T--Tianjin--cf-m3.png" alt="T--Tianjin--cf-m3" height="250px"> |
− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/7/75/T--Tianjin--cf-m4.png" alt="T--Tianjin--cf-m4" width="800" height="533"> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/7/75/T--Tianjin--cf-m4.png" alt="T--Tianjin--cf-m4" height="250px"> |
− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/b/be/T--Tianjin--cf-m5.png" alt="T--Tianjin--cf-m5" width="800" height="533"> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/b/be/T--Tianjin--cf-m5.png" alt="T--Tianjin--cf-m5" height="250px"> |
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− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/4/4e/T--Tianjin--cf-m7.png" alt="T--Tianjin--cf-m7" width="800" height="533"> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/4/4e/T--Tianjin--cf-m7.png" alt="T--Tianjin--cf-m7" height="250px"> |
− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/9/97/T--Tianjin--cf-m8.png" alt="T--Tianjin--cf-m8" width="800" height="533"> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/9/97/T--Tianjin--cf-m8.png" alt="T--Tianjin--cf-m8" height="250px"> |
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− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/c/c9/T--Tianjin--cf-m11.png" alt="T--Tianjin--cf-m11" width="800" height="533"> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/c/c9/T--Tianjin--cf-m11.png" alt="T--Tianjin--cf-m11" height="250px"> |
− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/3/36/T--Tianjin--cf-m12.png" alt="T--Tianjin--cf-m12" width="800" height="533"> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/3/36/T--Tianjin--cf-m12.png" alt="T--Tianjin--cf-m12"height="250px"> |
− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/d/d2/T--Tianjin--cf-m13.png" alt="T--Tianjin--cf-m13" width="800" height="533"> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/d/d2/T--Tianjin--cf-m13.png" alt="T--Tianjin--cf-m13" height="250px"> |
− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/3/35/T--Tianjin--cf-m14.png" alt="T--Tianjin--cf-m14" width="800" height="533"> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/3/35/T--Tianjin--cf-m14.png" alt="T--Tianjin--cf-m14" height="250px"> |
− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/b/b3/T--Tianjin--cf-m15.png" alt="T--Tianjin--cf-m15" width="800" height="533"> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/b/b3/T--Tianjin--cf-m15.png" alt="T--Tianjin--cf-m15" height="250px"> |
− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/2/2c/T--Tianjin--cf-m16.png" alt="T--Tianjin--cf-m16" width="800" height="533"> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/2/2c/T--Tianjin--cf-m16.png" alt="T--Tianjin--cf-m16" height="250px"> |
− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/9/91/T--Tianjin--cf-m17.png" alt="T--Tianjin--cf-m17" width="800" height="533"> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/9/91/T--Tianjin--cf-m17.png" alt="T--Tianjin--cf-m17" height="250px"> |
− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/3/3f/T--Tianjin--cf-m18.png" alt="T--Tianjin--cf-m18" width="800" height="533"> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/3/3f/T--Tianjin--cf-m18.png" alt="T--Tianjin--cf-m18" height="250px"> |
− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/a/ad/T--Tianjin--cf-m19.png" alt="T--Tianjin--cf-m19" width="800" height="533"> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/a/ad/T--Tianjin--cf-m19.png" alt="T--Tianjin--cf-m19" height="250px"> |
− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/0/03/T--Tianjin--cf-m20.png" alt="T--Tianjin--cf-m20" width="800" height="533"> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/0/03/T--Tianjin--cf-m20.png" alt="T--Tianjin--cf-m20" height="250px"> |
− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/b/b9/T--Tianjin--cf-m21.png" alt="T--Tianjin--cf-m21" width="800" height="533"> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/b/b9/T--Tianjin--cf-m21.png" alt="T--Tianjin--cf-m21" height="250px"> |
− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/3/3d/T--Tianjin--cf-m22.png" alt="T--Tianjin--cf-m22" width="800" height="533"> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/3/3d/T--Tianjin--cf-m22.png" alt="T--Tianjin--cf-m22" height="250px"> |
− | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/3/3e/T--Tianjin--cf-m-no.png" alt="T--Tianjin--cf-m-no" width="800" height="533"><br/> | + | <img class="col-md-4" src="https://static.igem.org/mediawiki/2016/3/3e/T--Tianjin--cf-m-no.png" alt="T--Tianjin--cf-m-no"height="250px"> |
| | | |
− | </div> | + | <br/> |
− | <a class="expand-btn">Show More</a> | + | |
| + | <p> |
| + | <b>The figures above show that the plasmids can express in the CFPS system, although the results of the parallel experimennts indicate that the CFPS system is unstable.</b> |
| + | </p> |
| + | |
| + | |
| + | |
| + | |
| + | </div> |
| + | <a class="expand-btn4">Show More</a> |
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− | <b>The figures above show that the plasmids can express in the CFPS system, although the results of the parallel experimennts indicate that the CFPS system is unstable.<b>
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| + | <!------------------------------------week5 start------------------------------------------------> |
| + | <div id="Week5"></div> |
| <article id="post-4252" class="post-4252 post type-post status-publish format-standard has-post-thumbnail hentry category-150 tag-174 tag-xinjiang tag-173"> | | <article id="post-4252" class="post-4252 post type-post status-publish format-standard has-post-thumbnail hentry category-150 tag-174 tag-xinjiang tag-173"> |
− | <h1 class="entry-title"><a href="https://www.camarts.cn/archives/4252.html" title="查看《火影忍者疾风传》中的全部作品" rel="bookmark">Week5(9/12/2016-9/18/2016)</a></h1> | + | <h1 class="entry-title">Week5(9/21/2016-9/27/2016)</h1> |
| <div class="entry-content"> | | <div class="entry-content"> |
− | <b>Ⅲ.Detection of Enzyme Activity<b> | + | |
| + | |
| + | <div class="note-content5"> |
| + | |
| + | <b>Ⅲ.Detection of Enzyme Activity</b> |
| <p> | | <p> |
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| </li> | | </li> |
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− | <li>Subsrtste: 1mM pNPA Tris-HCl buffer.<br/> | + | <li>Subsrtrate: 1mM pNPA Tris-HCl buffer.<br/> |
| 10 times diluted Enzyme solution(unpurified) mixed with the equal volume substrate.<br/> | | 10 times diluted Enzyme solution(unpurified) mixed with the equal volume substrate.<br/> |
| This attempt of detection failed, too. Because we found that Tris can react with pNPA , and pNPA would be transfered to pNP, too. | | This attempt of detection failed, too. Because we found that Tris can react with pNPA , and pNPA would be transfered to pNP, too. |
| </li></p> | | </li></p> |
− | <p> <a href="https://www.camarts.cn/archives/4252.html" class="more-link">查看Team Tianjin全部实验 <span class="meta-nav">›</span></a></p>
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− | <!-- #post-4252 --> | + | <!------------------------------------week5 end------------------------------------------------> |
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| + | |
| + | |
| + | <!------------------------------------week6 start------------------------------------------------> |
| + | <div id="Week6"></div> |
| + | <article id="post-4252" class="post-4252 post type-post status-publish format-standard has-post-thumbnail hentry category-150 tag-174 tag-xinjiang tag-173"> |
| <header class="entry-header"> | | <header class="entry-header"> |
− | <h1 class="entry-title"><a href="https://www.camarts.cn/archives/4252.html" title="查看《火影忍者疾风传》中的全部作品" rel="bookmark">Week6(9/19/2016-9/25/2016)</a></h1> | + | <h1 class="entry-title">Week6(9/28/2016-10/2/2016)</h1> |
| </header><!-- .entry-header --> | | </header><!-- .entry-header --> |
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| <div class="entry-content"> | | <div class="entry-content"> |
− | <p>
| + | |
| + | |
| + | <div class="note-content6"> |
| + | |
| + | <p> |
| <li>Substrate:0.2mM pNPA water solution.<br/> | | <li>Substrate:0.2mM pNPA water solution.<br/> |
| 10 times diluted Enzyme solution(unpurified) mixed with the equal volume substrate.<br/> | | 10 times diluted Enzyme solution(unpurified) mixed with the equal volume substrate.<br/> |
| After static reaction at 39℃ for 8 hours, we detected the characteristic adsorption peak of the product ,pNP, which has no other characteristic adsorption peak except in 400nm. | | After static reaction at 39℃ for 8 hours, we detected the characteristic adsorption peak of the product ,pNP, which has no other characteristic adsorption peak except in 400nm. |
− | </li>
| |
− | <li>Substrate:PET.<br/>
| |
− | PET was been put into 20 times diluted Enzyme solution(unpurified).<br/>
| |
− | After static reaction at 39℃ for 48 hours, we detected the characteristic adsorption peak of the product ,MHET, which has no other characteristic adsorption peak except in 260nm. Besides, we also deteected the system by Multispectral Scanning.
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| </li> | | </li> |
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− | <br /> | + | <img class="aligncenter size-full wp-image-4256" src="https://static.igem.org/mediawiki/2016/f/f4/T--Tianjin--cell-free_note-3.png" alt="T--Tianjin--cf-m-no" width="800" height="533"> |
| | | |
| + | <b>The control group in the image above indicate that something else in the CFPS system can cause characteristic adsorption peak except in 400nm with the sunstrate.</b><br/><br/><br/> |
| | | |
| + | <li>Substrate:PET.<br/> |
| + | PET was been put into 20 times diluted Enzyme solution(unpurified).<br/> |
| + | After static reaction at 39℃ for 5 days, we detected the characteristic adsorption peak of the product ,MHET, which has no other characteristic adsorption peak except in 260nm. Besides, we also deteected the product by Multispectral Scanning. |
| + | </li></p> |
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− | <p> <a href="https://www.camarts.cn/archives/4252.html" class="more-link">查看Team Tianjin全部实验 <span class="meta-nav">›</span></a></p>
| + | Please visit our Result part for more detailed result. |
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− | <h1 class="entry-title"><a href="https://www.camarts.cn/archives/4252.html" title="查看《火影忍者疾风传》中的全部作品" rel="bookmark">Week7(9/26/2016-10/2/2016)</a></h1>
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− | <p>
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− | <li>Plasmids pT-13-19-15 were handed in for sequencing, which confirmed its correctness.</li>
| |
− | <b>The sequencing results for them were correct.</b>
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− |
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− | <br />
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− | <b>Notice:</b> This page is currently under construction. Contents in this page are temporaory and will be modified several times before the final release. — 2016 iGEM Team Tianjin | + | <b>Notice: </b> This page is currently under construction. Contents in this page are temporaory and will be modified several times before the final release. — 2016 iGEM Team Tianjin |
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