Team:NTU Taiwan/index.html

     <title> Igem-Taiwan </title>

             <h1 class=" rainbow-text header">IGem-Taiwan Yeastherm</h1>

                 Working with Thermogenic Yeast<br/>

                 Apps with knowledge of iGEM competition and synthetic biology.

                 <p class="header" style="margin: 0"> our works </p>

 

                <p class="header" style="margin: 0"></p>

                 In hope of ?? the value of our biological heating device, we strive to improving the sensitivity of our sensor － the cold shock promoter. This final goal can be break down on two parts: 1. tuning the temperature-responsive range of cold shock promoter 2. amplifying its signal under low temperature. In order to understand what kind of structure of a genetic circuit and what kinds of characteristics of activator and repressor are needed for our purpose, we create several mathematical models to explore the problem. In the end, we expect to get some useful information as a guidance to screen possible biological parts when we actually start to construct the genetic circuit.

                             <td class="col-md-2">β_Csp(T)^a</td>

                             <td class="col-md-2">M/s, normalized</td>

         <br/><p><b>PCR</b></p><br/>

        &nbsp&nbsp&nbsp&nbsp&nbspStep 1: Design of appropriate forward and reverse primers<br/>

        &nbsp&nbsp&nbsp&nbsp&nbspStep 2: Prepare our template<br/>

        &nbsp&nbsp&nbsp&nbsp&nbspStep 3: Prepare the PCR mix. (Kapa Hifi PCR kit.)<br/>

        &nbsp&nbsp&nbsp&nbsp&nbspStep 4: Run PCR<br/>

        &nbsp&nbsp&nbsp&nbsp&nbspStep 5: Examine the results by electrophoresis<br/>

         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/><p><b>Construction of our parts</b></p><br/>

        &nbsp&nbsp&nbsp&nbsp&nbspStep 1: We design primers for parts with prefix and suffix.<br/>

        &nbsp&nbsp&nbsp&nbsp&nbspStep 2: Perform PCR and cleanup the PCR product<br/>

        &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/>

        &nbsp&nbsp&nbsp&nbsp&nbspStep 4: Ligation of inserts and backbones<br/>

        &nbsp&nbsp&nbsp&nbsp&nbspStep 5: Transform our ligation products into DH5α and streak the transformed DH5α on LB agar plate with chloramphenicol.<br/>

        &nbsp&nbsp&nbsp&nbsp&nbspStep 6: Inoculate single colony into broth with chloramphenicol.<br/>

        &nbsp&nbsp&nbsp&nbsp&nbspStep 7: Miniprep the plasmid DNA from the overnight broth culture.<br/>

        &nbsp&nbsp&nbsp&nbsp&nbspStep 8: Confirm the products by both RE digestion and PCR sequencing<br/>

                 <p>However, we didn’t analyze the quantity of yeasts during the experiment. We consider that the experimental group (which had been transformed SrUCP) might grow slower than the control group and then cause the result t I show below:

                     Experimental: Heat(E) = Fermentation(1) + SrUCP

                     Control: Heat(C) = Fermentation(2)

                 <img src="https://www.dropbox.com/s/wtpek4rg4x1s8vt/Capture2.JPG" alt-src="images/Capture2.jpg">

                 <img src="https://www.dropbox.com/s/wtpek4rg4x1s8vt/Capture2.JPG" alt-src="images/Capture3.jpg">