Team:NTU Taiwan/index.html

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     <title> Igem-Taiwan </title>
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     <title> iGEM-NTU-Taiwan </title>
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             <h1 class=" rainbow-text header">IGem-Taiwan Yeastherm</h1>
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             <h1 class=" rainbow-text header">iGEM-NTU-Taiwan YeasTherm</h1>
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                 <img class="spin" alt-src="images/LaboratoryLevels.png" src="/wiki/images/9/91/NTU_TAIWAN_LaboratoryLevels.png"><br/>
                 <img class="spin" alt-src="images/LaboratoryLevels.png" src="/wiki/images/9/91/NTU_TAIWAN_LaboratoryLevels.png"><br/>
                 National Taiwan University<br/>
                 National Taiwan University<br/>
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                 Working with Thermogenic Yeast<br/>
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                 Working on Thermogenic Yeast<br/>
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                 Apps with knowledge of iGEM competition and synthetic biology.
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                 Apps with concept of iGEM competition and synthetic biology.
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         <br/><p><b>PCR</b></p><br/><p>
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         <br/><p><b>PCR</b></p>
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        &nbsp&nbsp&nbsp&nbsp&nbspStep 1: Design of appropriate forward and reverse primers<br/>
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1: Design of appropriate forward and reverse primers<br/>
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        &nbsp&nbsp&nbsp&nbsp&nbspStep 2: Prepare our template<br/>
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2: Prepare our template<br/>
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        &nbsp&nbsp&nbsp&nbsp&nbspStep 3: Prepare the PCR mix. (Kapa Hifi PCR kit.)<br/>
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3: Prepare the PCR mix. (Kapa Hifi PCR kit.)<br/>
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        &nbsp&nbsp&nbsp&nbsp&nbspStep 4: Run PCR<br/>
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4: Run PCR<br/>
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        &nbsp&nbsp&nbsp&nbsp&nbspStep 5: Examine the results by electrophoresis<br/>
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5: Examine the results by electrophoresis<br/>
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         Note: If the template is genomic DNA, we would adjust the annealing temperature at 45°C. It is because the copy number of target gene may be low. We use this annealing temp when perform PCR of Tir1, 26s, 5.8s ITS</p>
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         Note: If the template is genomic DNA, we would adjust the annealing temperature at 45°C. It is because the copy number of target gene may be low. We use this annealing temp when perform PCR of Tir1, 26s, 5.8s ITS<br/><br/>
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<br/><p><b>Construction of our parts</b></p><br/><p>
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<br/><p><b>Construction of our parts</b></p><p>
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        &nbsp&nbsp&nbsp&nbsp&nbspStep 1: We design primers for parts with prefix and suffix.<br/>
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1: We design primers for parts with prefix and suffix.<br/>
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        &nbsp&nbsp&nbsp&nbsp&nbspStep 2: Perform PCR and cleanup the PCR product<br/>
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2: Perform PCR and cleanup the PCR product<br/>
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        &nbsp&nbsp&nbsp&nbsp&nbspStep 3: Before insert our parts into standard backbone, pSB1C3, we perform RE digestion to make sticky ends of both inserts and backbones.<br/>
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3: Before insert our parts into standard backbone, pSB1C3, we perform RE digestion to make sticky ends of both inserts and backbones.<br/>
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        &nbsp&nbsp&nbsp&nbsp&nbspStep 4: Ligation of inserts and backbones<br/>
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4: Ligation of inserts and backbones<br/>
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        &nbsp&nbsp&nbsp&nbsp&nbspStep 5: Transform our ligation products into DH5α and streak the transformed DH5α on LB agar plate with chloramphenicol.<br/>
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5: Transform our ligation products into DH5α and streak the transformed DH5α on LB agar plate with chloramphenicol.<br/>
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        &nbsp&nbsp&nbsp&nbsp&nbspStep 6: Inoculate single colony into broth with chloramphenicol.<br/>
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6: Inoculate single colony into broth with chloramphenicol.<br/>
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        &nbsp&nbsp&nbsp&nbsp&nbspStep 7: Miniprep the plasmid DNA from the overnight broth culture.<br/>
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7: Miniprep the plasmid DNA from the overnight broth culture.<br/>
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        &nbsp&nbsp&nbsp&nbsp&nbspStep 8: Confirm the products by both RE digestion and PCR sequencing<br/></p>
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8: Confirm the products by both RE digestion and PCR sequencing<br/></p>
         <p><b>Point mutation protocol</b></p>
         <p><b>Point mutation protocol</b></p>

Latest revision as of 04:22, 28 September 2013

iGEM-NTU-Taiwan

Retrieved from "http://2013.igem.org/Team:NTU_Taiwan/index.html"