Team:DTU-Denmarkgogo

From 2014.igem.org

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<pageheader>Meet the team</pageheader>
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<pageheader>Protocols</pageheader>
<regulartext>
<regulartext>
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<tr>
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<th colspan="2" scope="col"><h3>Glycerol stock-preservation</h3></th>
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<th colspan="2" scope="col"><h1>Glycerol stock-preservation</h1></th>
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<tr>
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<td>
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<b>Motivation</b><br>
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I joined our team to experience how to manage a scientific project from the very beginning and to the end, to improve my team working skills as well as to work with some crazy science. Of course being shielded from the sun by spending most of my waking hours in front of a lab bench during the summer also attracted.
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<ol>
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<br><br>
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<li>Add one ml sterile 50% glycerol to a cryotube.</li>
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<b>Major contributions</b><br>
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<li>Add one ml overnight culture to a cryotube from step 1</li>
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Coordination of the fundraising and sponsorships as well as maintaining sterile technique i the lab.
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<li>Store at -80 °C</li>
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<br><br>
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</ol>
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<b>How do you prefer spinach?</b><br>
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Try frozen spinach on a pizza with onion, goat cheese and a lot of garlic - that is delicious.
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<th colspan="2" scope="col"><h2>Caroline Mosbech</h2></th>
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<th colspan="2" scope="col"><h1>PCR</h1></th>
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<tr>
<tr>
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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="180"/> </a> </center>
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<tr>
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<td>
<td>
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bla<br>
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bla<br>
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bla<br>
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<ul>
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bla<br>
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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>
-
bla<br>
+
<li>The polymerase stock must only be taken out of the freezer when needed and must be put back immediately after use.</li>
-
bla<br>
+
<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>
 +
</ul>
 +
 
 +
 
</td>
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</tr>
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</table>
 
<table width="100%" border="0" cellspacing="15" cellpadding="0" style="background-color:transparent;">
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<tr>
<tr>
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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>
</tr>     
</tr>     
<tr>
<tr>
<td>
<td>
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<b>Motivation</b><br>
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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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<ol>
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<b>Major contributions</b><br>
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<li>A</li>
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Wiki geek<br>
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<li>A</li>
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                                  <br>
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<li>S</li>
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<b>How do you prefer spinach?</b><br>
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</ol>
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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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<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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<tr>
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<td>
<td>
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bla<br>
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bla<br>
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bla<br>
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<ol>
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bla<br>
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<li>h</li>
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bla<br>
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<li>u</li>
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<li>i</li>
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</ol>
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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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<b>Motivation</b><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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<b>Major contributions</b><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>
 +
            <li>2.5g NaCl</li>
 +
            <li>5.0g NH4Cl</li>
 +
            <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>
 +
                      </ul>
 +
<li>Measure ~700ml of distilled H2O (sterile)</li>
 +
<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>
 +
 
 +
</ol>
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</td>
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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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<td>
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<b>Motivation</b><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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<b>Major contributions</b><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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<b>How do you prefer spinach?</b><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>
 +
<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>
 +
<li>Analyze your gel</li>
 +
</ol>
 +
 
</td>
</td>
</tr>
</tr>
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</table>
</table>
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<table width="100%" border="0" cellspacing="15" cellpadding="0" style="background-color:transparent;">
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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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<td>
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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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                      </ul>
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<ol>
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<li></li>
 +
<li></li>
 +
<li></li>
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<li></li>
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</ol>
 +
 
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<b>Motivation</b><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>
 
-
<br>
 
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<b>Major contributions</b><br>
 
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<br>
 
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<b>How do you prefer spinach?</b><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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<td rowspan="1" scope="col" width="300px">
<td rowspan="1" scope="col" width="300px">
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<img style="float: left; "src="https://static.igem.org/mediawiki/2014/2/23/Thor.jpg" height="300"> </img>
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<img style="float: left; "src="http://i.perezhilton.com/wp-content/uploads/2010/03/popeye46704__oPt.jpg" height="300"> </img>
</td>
</td>
<td>
<td>
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<b>Motivation</b><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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<b>Major contributions</b><br>
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-
<br>
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-
<br>
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<b>How do you prefer spinach?</b><br>
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Luminescent and far away from my taste buds.<br>
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<img style="float: left; "src="https://static.igem.org/mediawiki/2014/4/47/Anne_Sofie.jpg" height="300"> </img>
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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

Retrieved from "http://2014.igem.org/Team:DTU-Denmarkgogo"