Team:DTU-Denmarkgogo

From 2014.igem.org

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<pageheader>Protocols</pageheader>
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<li> Add one ml sterile 50% glycerol to a cryotube<li/>
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<li>Add one ml sterile 50% glycerol to a cryotube.</li>
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<li>Add one ml overnight culture to a cryotube from step 1<li/>
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<li>Add one ml overnight culture to a cryotube from step 1</li>
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<li>Store at -80 °C<li/>
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<li>Store at -80 °C</li>
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<th colspan="2" scope="col"><h1>PCR</h1></th>
<th colspan="2" scope="col"><h1>PCR</h1></th>
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<center><a href="h"><img src="https://static.igem.org/mediawiki/2014/8/8e/PCR_PIC.png" width="942" height="180"/> </a> </center>
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<li>Make a mastermix of the reagents that are common for all reactions. Make enough for 0,5 extra reaction. Add reagents in the order they are listed above.</li>
 +
<li>The polymerase stock must only be taken out of the freezer when needed and must be put back immediately after use.</li>
 +
<li>Remember to mix thoroughly before aliquotting to separate PCR tubes, and try not to make any bubbles. Reverse pipetting works well for aliquoting.</li>
 +
<li>Mark the PCR tubes on lid and side and place in thermocycler.</li>
 +
<li>The annealing temperature should match the primer melting temperatures. This can be hard as Tm predictors tend to disagree.</li>
 +
<li>Elongation time should be AT LEAST 30 seconds per kb of the longest expected fragment (for the X7 polymerase).</li>
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</ul>
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<td>
 
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<center><a  href="h"><img src="https://static.igem.org/mediawiki/2014/8/8e/PCR_PIC.png" width="942" height="200"/>
 
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</a>
 
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bla<br>
 
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<th colspan="2" scope="col"><h2>Georgi Dimitrov</h2></th>
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<th colspan="2" scope="col"><h1>Casting of gels and gel electrophoresis (remember to include EtBr)</h1></th>
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<tr>
<tr>
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Motivation <br>
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<td>
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My motivation is to gain as much experience as possible - Wiki page, Lab work ,Human relations and have a Crazy summer .My priority is to learn a lot about the practical part of Systems biology and to contribute to the  science community  if possible  ,specially to all biohachers and citizen scientists all over the worlds.<br>
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                                  <br>
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Major contributions<br>
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<ol>
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Wiki geek<br>
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<li>A</li>
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                                  <br>
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<li>A</li>
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How do you prefer spinich?<br>
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<li>S</li>
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The Balkan way. I go camping somewhere and I make soup out of it and I put it to cook very slowly on a    big fire in a pot.When the soap is done I throw it out and I dig out the roast lamb from under the fire.<br>
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</ol>
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<th colspan="2" scope="col"><h2>Kristian Barret Karlsen</h2></th>
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<th colspan="2" scope="col"><h1>Preparation of Cam stock solution</h1></th>
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<th colspan="2" scope="col"><h2>Kristian Jensen</h2></th>
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<th colspan="2" scope="col"><h1>Recipe on M9 minimal media</h1></th>
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Motivation<br>
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I am very interested in synthetic biology, so joining the university’s iGEM team seemed like an obvious choice. I am excited about the prospect of completing our own research project and hopefully contributing to the synthetic biology community.<br>
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<ol>
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<br>
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<li>Make M9 salts</li>
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Major contributions<br>
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<li>To make M9 Salts aliquot 800ml H2O and add</li>
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<br>
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        <ul>
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How do you prefer spinach?<br>
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            <li>64g Na2HPO4-7H2O</li>
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Juiced - with a dash of vodka (or two).<br>
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            <li>15g KH2PO4</li>
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            <li>2.5g NaCl</li>
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            <li>5.0g NH4Cl</li>
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            <li>Stir until dissolved</li>
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            <li>Adjust to 1000ml with distilled H2O</li>
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            <li>Sterilize by autoclaving</li>
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                      </ul>
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<li>Measure ~700ml of distilled H2O (sterile)</li>
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<li>Add 200ml of M9 salts</li>
 +
<li>Add 2ml of 1M MgSO4 (sterile)</li>
 +
<li>Add 20 ml of 20% glucose (or other carbon source)</li>
 +
<li>Add 100ul of 1M CaCl2 (sterile)</li>
 +
<li>Adjust to 1000ml with distilled H2O</li>
 +
 
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</ol>
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<th colspan="2" scope="col"><h2>Mark Thomas Østerlund</h2></th>
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<th colspan="2" scope="col"><h1>Restriction analysis</h1></th>
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Motivation<br>
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I have a deep fascination with the possibilities of synthetic biology. The fact that we are know capable of tailoring organisms to fit our specific needs is just astonishing! Besides it’s a great opportunity to pick up some new skills relevant to working in a different kind of group.<br>
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<ol>
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<br>
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<li>Make a mastermix containing.</li>
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Major contributions<br>
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  <ul>
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Wiki<br>
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            <li>1X NEBuffer (according to the desired restriction enzyme)</li>
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<br>
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            <li>1X BSA (note if NEBuffer is called X.1 BSA is already added in the buffer)</li>
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How do you prefer spinach?<br>
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            <li>1 unit/μl restriction enzyme</li>
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In a salmon-spinach pie.<br>
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                      </ul>
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<li>Divide the mastermix into tubes and add DNA to a final concentration of 10 ng/μl</li>
 +
<li>Incubate the reaction mix at the restriction enzymes optimal temperature for 30-60 minutes</li>
 +
<li>Add loading buffer to samples and run on 1% agarose gel with ethidium bromide 0.2 μg/ml</li>
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<li>Analyze your gel</li>
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</ol>
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<th colspan="2" scope="col"><h2>Rasmus Bech</h2></th>
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<th colspan="2" scope="col"><h1>next</h1></th>
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                      </ul>
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<li></li>
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<li></li>
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<li></li>
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<li></li>
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</ol>
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<td>
 
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Motivation<br>
 
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I joined iGEM in order to take part in a project where the students have more control and responsibility than other projects at DTU. It was also a motivation being part of a team, as large projects at DTU most often are individual.<br>
 
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<br>
 
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Major contributions<br>
 
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<br>
 
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How do you prefer spinach?<br>
 
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I prefer spinach in a salad with strawberry, red onion, pine nuts, feta and a dressing with Balsamico vinegar, oil, sugar, soy sauce and tabasco.<br>
 
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<th colspan="2" scope="col"><h2>Thor Bech Johannesen</h2></th>
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<th colspan="2" scope="col"><h1>Protocol for eating spinach</h1></th>
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<img style="float: left; "src="http://i.perezhilton.com/wp-content/uploads/2010/03/popeye46704__oPt.jpg" height="300"> </img>
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</td>
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Motivation<br>
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As a student at DTU you get to do plenty of project work, however it is mostly carried out individually or in a small group, with very specific goals and a high degree of supervision. iGEM offers the opportunity to work on a much larger project with a high degree of autonomy, where everything from the overall goals and strategy to design of experiments is left for the students to manage. This provides a great opportunity to experience the challenges of actual research projects<br>
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<br>
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Major contributions<br>
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<br>
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<br>
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How do you prefer spinach?<br>
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Luminescent and far away from my taste buds.<br>
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</td>
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Latest revision as of 14:32, 1 August 2014

Protocols

Glycerol stock-preservation
  1. Add one ml sterile 50% glycerol to a cryotube.
  2. Add one ml overnight culture to a cryotube from step 1
  3. Store at -80 °C

PCR
  • Make a mastermix of the reagents that are common for all reactions. Make enough for 0,5 extra reaction. Add reagents in the order they are listed above.
  • The polymerase stock must only be taken out of the freezer when needed and must be put back immediately after use.
  • Remember to mix thoroughly before aliquotting to separate PCR tubes, and try not to make any bubbles. Reverse pipetting works well for aliquoting.
  • Mark the PCR tubes on lid and side and place in thermocycler.
  • The annealing temperature should match the primer melting temperatures. This can be hard as Tm predictors tend to disagree.
  • Elongation time should be AT LEAST 30 seconds per kb of the longest expected fragment (for the X7 polymerase).

Casting of gels and gel electrophoresis (remember to include EtBr)
  1. A
  2. A
  3. S

Preparation of Cam stock solution
  1. h
  2. u
  3. i

Recipe on M9 minimal media
  1. Make M9 salts
  2. To make M9 Salts aliquot 800ml H2O and add
    • 64g Na2HPO4-7H2O
    • 15g KH2PO4
    • 2.5g NaCl
    • 5.0g NH4Cl
    • Stir until dissolved
    • Adjust to 1000ml with distilled H2O
    • Sterilize by autoclaving
  3. Measure ~700ml of distilled H2O (sterile)
  4. Add 200ml of M9 salts
  5. Add 2ml of 1M MgSO4 (sterile)
  6. Add 20 ml of 20% glucose (or other carbon source)
  7. Add 100ul of 1M CaCl2 (sterile)
  8. Adjust to 1000ml with distilled H2O

Restriction analysis
  1. Make a mastermix containing.
    • 1X NEBuffer (according to the desired restriction enzyme)
    • 1X BSA (note if NEBuffer is called X.1 BSA is already added in the buffer)
    • 1 unit/μl restriction enzyme
  2. Divide the mastermix into tubes and add DNA to a final concentration of 10 ng/μl
  3. Incubate the reaction mix at the restriction enzymes optimal temperature for 30-60 minutes
  4. Add loading buffer to samples and run on 1% agarose gel with ethidium bromide 0.2 μg/ml
  5. Analyze your gel

next

Protocol for eating spinach

Overview

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