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| <span>1.Photos of gel electrophoresis (confirming the successful construction of all gene circuit) </span> | | <span>1.Photos of gel electrophoresis (confirming the successful construction of all gene circuit) </span> |
| <span><a href="javascript:return false;" onClick="route(2)">click here</a></span> | | <span><a href="javascript:return false;" onClick="route(2)">click here</a></span> |
− | <span>2. CI binding site insertion with whole plasmid PCR</span> | + | <span>2. Experiments proving the efficiency of the antimicrobial peptide</span> |
| <span><a>click here</a></span> | | <span><a>click here</a></span> |
− | <span>3. Experiments proving the efficiency of the antimicrobial peptide</span> | + | <span>3. CI binding site insertion with whole plasmid PCR</span> |
| <span><a>click here</a></span> | | <span><a>click here</a></span> |
| </div> | | </div> |
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| </div> | | </div> |
− | <div class="editarea" id="member8">
| + | </div> |
− | <h2 style="display:inline;float:left">CI Binding Site Insertion_A Brand-new PCR Strategy</h2>
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− | <h2 style="display:inline;float:right">Page 1 of 7</h2>
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− | <div class="member">
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− | <span>Inspired by the site-directed mutagenesis and the overlap PCR, we developed a two-step whole plasmid PCR strategy that enables precise insertion of a relatively long DNA segment in any site of interest. For the great convenience this strategy could bring, we decided to show it in our result. Advantages are listed below:</span>
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− | <span>1.It saves a lot of time from molecular cloning. Teams of iGEM always spend a lot of time on the digestion with restriction endonuclease to obtain the target DNA and the ligation to construct a composite part. Also, adding a tag to a basic part using ordinary overlap PCR requires the subsequent reconstruction of all the composite parts containing that basic part. However, with the assistance of our PCR strategy, teams are able to get a whole plasmid ready to be transformed after the insertion at a predefined site.</span>
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− | <span>2. It enables the insertion of a relatively long DNA segment. The site-directed mutagenesis could be used for insertion in a whole plasmid, but the insertion is merely several base pairs long. Our PCR strategy is proved to insert a DNA segment (i.e. the 46-bp long CI binding site) which is longer than what the site-directed mutagenesis could provide. Theoretically speaking, it could even afford insertion of longer DNA segments. </span>
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− | <span>We successfully inserted the CI binding site into the specific site after failed many times with the ordinary assembly strategy.</span>
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− | <span>Step1:</br>The first pair of primers are adjacent to each other with opposite direction. After amplification by PCR, circular plasmids are “opened” at a specific cite of interest and linear plasmids are obtained (see Fig.1). </span>
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− | <img src="https://static.igem.org/mediawiki/2015/5/5a/WHU-China_ResultPart2_1.jpg" height="200">
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− | <span style="margin-top:210px">Fig.1 The first pair of primers "open" the circular plasmid at a specific cite. </span>
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− |
| + | |
− | </div>
| + | |
− | </div>
| + | |
− | <div class="editarea" id="member9">
| + | |
− | <h2 style="display:inline;float:left">CI Binding Site Insertion_A Brand-new PCR Strategy</h2>
| + | |
− | <h2 style="display:inline;float:right">Page 1 of 7</h2>
| + | |
− | <div class="member">
| + | |
− | <span>Inspired by the site-directed mutagenesis and the overlap PCR, we developed a two-step whole plasmid PCR strategy that enables precise insertion of a relatively long DNA segment in any site of interest. For the great convenience this strategy could bring, we decided to show it in our result. Advantages are listed below:</span>
| + | |
− | <span>1.It saves a lot of time from molecular cloning. Teams of iGEM always spend a lot of time on the digestion with restriction endonuclease to obtain the target DNA and the ligation to construct a composite part. Also, adding a tag to a basic part using ordinary overlap PCR requires the subsequent reconstruction of all the composite parts containing that basic part. However, with the assistance of our PCR strategy, teams are able to get a whole plasmid ready to be transformed after the insertion at a predefined site.</span>
| + | |
− | <span>2. It enables the insertion of a relatively long DNA segment. The site-directed mutagenesis could be used for insertion in a whole plasmid, but the insertion is merely several base pairs long. Our PCR strategy is proved to insert a DNA segment (i.e. the 46-bp long CI binding site) which is longer than what the site-directed mutagenesis could provide. Theoretically speaking, it could even afford insertion of longer DNA segments. </span>
| + | |
− | <span>We successfully inserted the CI binding site into the specific site after failed many times with the ordinary assembly strategy.</span>
| + | |
− | <span>Step1:</br>The first pair of primers are adjacent to each other with opposite direction. After amplification by PCR, circular plasmids are “opened” at a specific cite of interest and linear plasmids are obtained (see Fig.1). </span>
| + | |
− | <img src="https://static.igem.org/mediawiki/2015/5/5a/WHU-China_ResultPart2_1.jpg" height="200">
| + | |
− | <span style="margin-top:210px">Fig.1 The first pair of primers "open" the circular plasmid at a specific cite. </span>
| + | |
− |
| + | |
− | </div>
| + | |
− | </div>
| + | |
− |
| + | |
| </div> | | </div> |
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