Difference between revisions of "Team:Kyoto/Safety"

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<h1> Safety </h1>
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<p>Please visit the <a href="https://2018.igem.org/Safety">Safety Hub</a> to find this year's safety requirements & deadlines, and to learn about safe & responsible research in iGEM.</p>
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<p>On this page of your wiki, you should write about how you are addressing any safety issues in your project. The wiki is a place where you can <strong>go beyond the questions on the safety forms</strong>, and write about whatever safety topics are most interesting in your project. (You do not need to copy your safety forms onto this wiki page.)</p>
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  <div id="jump"><a href="#wrapper"><img src="https://static.igem.org/mediawiki/2017/c/c5/Kyoto_notebook_jump.png"></a></div>
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  <div id="BACKGROUND">
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  <h1>Material&amp;Methods</h1>
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  <ul class="material">
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      <li><a href="#Parts">1) Parts</a></li>
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      <li><a href="#Primer">2) Primer list</a></li>
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      <li><a href="#Materials">3) Materials</a></li>
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        <ul>
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<li><a href="#Kit">3-1 Kit</a></li>
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<li><a href="#Restriction Enzyme">3-2 Restriction Enzyme</a></li>
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<li><a href="#Polymerase">3-3 Polymerase</a></li>
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<li><a href="#DNA ligase">3-4 DNA ligase</a></li>
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<li><a href="#Marker">3-5 Marker</a></li>
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<li><a href="#Organism">3-6 Organism</a></li>
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<li><a href="#Antibiotics">3-7 Antibiotics</a></li>
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<li><a href="#Equipment">3-8 Equipment</a></li>
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<li><a href="#Backbone">3-9 Backbone</a></li>
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<li><a href="#Buffer">3-10 Buffer</a></li>
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<li><a href="#SDS-PAGE&Westernblotting">3-11 SDS-PAGE&Westernblotting</a></li>
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<!--------------------------------------------------------- BLOCK Contents-2 Materials  END----------------------------------------------------------->
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        </ul>
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  <li><a href="#methods">4) Methods</a></li>
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    <ul>
  
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<!--------------------------------------------------------- BLOCK Contents-3 Methods PY------------------------------------------------------------>
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<li><a href="#Miniprep">4-1 Miniprep</a></li>
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<li><a href="#Gel Extraction and PCR purification">4-2 Gel Extraction and PCR purification</a></li>
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<li><a href="#Restriction Enzyme Digestion">4-3 Restriction Enzyme Digestion</a></li>
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<li><a href="#Ligation">4-4 Ligation</a></li>
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<li><a href="#Transformation">4-5 Transformation</a></li>
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<!--------------------------------------------------------- BLOCK Contents-3 Methods  END------------------------------------------------------------>
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  </ul>
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</ul>
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<!-- ######################################################  BLOCK Contents END #################################################-->
  
  
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<div class="column two_thirds_size">
 
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<h3>Safe Project Design</h3>
 
  
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<p>Does your project include any safety features? Have you made certain decisions about the design to reduce risks? Write about them here! For example:</p>
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<!-----------------------------------------------------BLOCK Parts-------------------------------------------------------------->
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<h5 id="Parts">1) Parts</h5>
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<h6><a href="https://2017.igem.org/Team:Kyoto/Basic_Part" target="brank">Basic Parts</a></h6>
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<h6><a href="https://2017.igem.org/Team:Kyoto/Composite_Part" target="brank">Composite Parts</a></h6>
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<!-----------------------------------------------------BLOCK Parts END---------------------------------------------------------->
  
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<ul>
 
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<li>Choosing a non-pathogenic chassis</li>
 
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<li>Choosing parts that will not harm humans / animals / plants</li>
 
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<li>Substituting safer materials for dangerous materials in a proof-of-concept experiment</li>
 
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<li>Including an "induced lethality" or "kill-switch" device</li>
 
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</ul>
 
  
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<!------------------------------------------------------BLOCK primer PY---------------------------------------------------------->
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<h3>Safe Lab Work</h3>
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<h5 id = "Primer">2) Primer List</h5>
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<tr><th>primer name</th><th class="example" width=40%>sequence</th><th width=8%>Length</th><th width=5%>Tm</th><th width=8%>GC%</th><th width=10%>Designer</th><th>Manufacturer</th></tr><tr><td>tropomyosinF</td><td>GATCGAGAAGGACAACGCCC</td><td>20</td><td>65</td><td>60</td><td>Fukuda</td><td>Macrogen Japan Corp</td></tr><tr><td>IK107</td><td>cccgccgccaccatggaggcggccgcaaaatcagg</td><td>35</td><td>94</td><td>71</td><td>Yoshimoto</td><td>Macrogen Japan Corp</td></tr></table>
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<h5 id="Materials">3) Materials</h5>
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<h6 id = "Kit">3-1 Kit</h6>
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<table>
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<tr><th>Name</th><th>Supplier</th></tr><tr><td>Wizard&#174; SV Gel and PCR</td><td>Promega</td></tr><tr><td>FastGene™Plasmid Mini Kit</td><td>NIPPON Genetics Co.,Ltd</td></tr>
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<h5 id="methods">4) Methods</h5>
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<h6 id = "Miniprep">4-1 Miniprep</h6>
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<p>
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Minipreps were performed using FastGene&#8482;Plasmid Mini Kit and and Wizard® Plus SV Minipreps DNA Purification System according to the manufacturer's protocols.
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<h6 id = "Soaking">4-17 Soaking</h6>
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<p>We followed this paper as for soaking.<br>
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  <a href="https://link.springer.com/content/pdf/10.1007%2Fs10658-012-0035-0.pdf">https://link.springer.com/content/pdf/10.1007%2Fs10658-012-0035-0.pdf</a></p>
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<p>What safety procedures do you use every day in the lab? Did you perform any unusual experiments, or face any unusual safety issues? Write about them here!</p>
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<h3>Safe Shipment</h3>
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<p>Did you face any safety problems in sending your DNA parts to the Registry? How did you solve those problems?</p>
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Revision as of 08:02, 2 October 2018

Material&Methods

1) Parts
Basic Parts
Composite Parts
2) Primer List
primer namesequenceLengthTmGC%DesignerManufacturer
tropomyosinFGATCGAGAAGGACAACGCCC206560FukudaMacrogen Japan Corp
IK107cccgccgccaccatggaggcggccgcaaaatcagg359471YoshimotoMacrogen Japan Corp
3) Materials
3-1 Kit
NameSupplier
Wizard® SV Gel and PCRPromega
FastGene™Plasmid Mini KitNIPPON Genetics Co.,Ltd
4) Methods
4-1 Miniprep

Minipreps were performed using FastGene™Plasmid Mini Kit and and Wizard® Plus SV Minipreps DNA Purification System according to the manufacturer's protocols.

4-17 Soaking

We followed this paper as for soaking.
https://link.springer.com/content/pdf/10.1007%2Fs10658-012-0035-0.pdf