Difference between revisions of "Team:KoreaSonyeodul/Experiments"

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<p>Describe the experiments, research and protocols you used in your iGEM project.</p>
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<h5>What should this page contain?</h5>
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<ul>
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<li> Protocols </li>
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<li> Experiments </li>
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<li>Documentation of the development of your project </li>
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</ul>
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<h5>Inspiration</h5>
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<ul>
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<li><a href="https://2014.igem.org/Team:Colombia/Protocols">2014 Colombia </a></li>
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<li><a href="https://2014.igem.org/Team:Imperial/Protocols">2014 Imperial </a></li>
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<li><a href="https://2014.igem.org/Team:Caltech/Project/Experiments">2014 Caltech </a></li>
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</ul>
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          <h1>
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            <font face="Roboto" weight="200">Experiments</font>
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          </h1>
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          <h2>
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            <font face="Roboto">Main Subtitle</font>
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          </h2>
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        <div class="Protocol1">
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          <h1>Protocol</h1>
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          <ul style="list-style-type: decimal;">
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            <li>Mealworm Dissection</li>
 +
            <li>
 +
              LB Agar Medium Synthesis
 +
              <h3>Materials Needed:</h3>
 +
              <ul style="list-style-type: disc;">
 +
                <li>Water(purified water)</li>
 +
                <li>
 +
                  LB broth (salt, yeast, 카세린), since we’re making a 500ml solution, we’ll need 12.5g
 +
                  <ul style="list-style-type: disc;">
 +
                    <li>25g/liter</li>
 +
                  </ul>
 +
                </li>
 +
                <li>
 +
                  AGAR (not easily disintegrated by bacteria, structure preserved)
 +
                  <ul style="list-style-type: disc;">
 +
                    <li>Has to be 1.5% of the solution</li>
 +
                    <li>Since we’re making a 500ml solution, we’ll need 7.5g</li>
 +
                  </ul>
 +
                </li>
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                <li>
 +
                  Autoclave
 +
                  <ul style="list-style-type: disc;">
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                    <li>
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                      High pressure, high temp  sterilization
 +
                    </li>
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                    <li>
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                      More than 20 minutes
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                    </li>
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                  </ul>
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                </ul>
 +
 +
              <h3>Procedure:</h3>
 +
              <ul style="list-style-type: lower-latin;">
 +
                <li>
 +
                  Put water up to 500ml in a graduated cylinder  pour it in a large flask
 +
                </li>
 +
                <li>Measure LB broth carefully to match 12.5g inside the a plastic temporary container</li>
 +
                <li>
 +
                  Close the lid to the LB broth, wash the spoon used to transfer it
 +
                </li>
 +
                <li>Fold the plastic container in half and pour the LB broth into the flask</li>
 +
                <li>In the same manner, put AGAR(7.5g) inside the flask </li>
 +
                <li>
 +
                  Don’t mix he flask, and seal the top of the flask with a tin foil
 +
                </li>
 +
                <li>Put it in the autoclave, pour some hot water and close the lid  wait for an hour and half</li>
 +
                <li>When storing the AGAR medium, make sure to preserve it in 60 degree cel.</li>
 +
                <li>Go to the clean bench, or if it’s unavailable you kindle the alcohol lamp(which causes convection and so in a sense forms an umbrella that protects the plate from contamination from the air) </li>
 +
                <li>
 +
                  Pour the AGAR solution onto eight plates, making sure your hand or your other body part is not hovering over the plate (you always have to keep in mind that SOMETHING’s falling from the air.
 +
                </li>
 +
                <li>
 +
                  Close the lids of the all eight plates and wait till they harden.<br/>
 +
                </li>
 +
 +
              </ul>
 +
 +
            </li>
 +
            </li>
 +
            <li>
 +
              Chemical Transformation
 +
              <ul style="none">
 +
                <li>
 +
                  Basic Knowledge:
 +
                  <ul style="list-style-type: circle;">
 +
                    <li>Competent cell</li>
 +
                    <li>e-coli’s name : DH5-alpha</li>
 +
                    <li>DNA to be Added : PUC19</li>
 +
                  </ul>
 +
                </li>
 +
              </ul>
 +
              <h3>Procedure:</h3>
 +
              <ul style="list-style-type: lower-latin;">
 +
                <li>Prepare two containers</li>
 +
                <ul style="list-style-type: lower-roman;">
 +
                  <li>only the DH5-alpha</li>
 +
                  <li>DH5 alpha + PUC19 (experimental group)</li>
 +
                </ul>
 +
                <li>Put both containers in ice for a minute</li>
 +
                <li>Raise the temperature abruptly to 42 degrees Cel for a minute</li>
 +
                <li>Put them into ice for another 2 minutes</li>
 +
                <li>At the Clean Bench, put LB Medium equally to each container</li>
 +
                <li>37&deg;C 180 RPM (at least 30-min)</li>
 +
                <li>In the centrifuge, 30 seconds at highest speed</li>
 +
                <li>With a Pipet eliminate remaining precipitate</li>
 +
                <li>In the LB Medium plate, put appropriate number of glass beads</li>
 +
                <li>After mixing the substances in the two containers with the pipet,</li>
 +
                <li>Put the two substances into each separate LB plates, put the lid on top and shake</li>
 +
                <li>Seal the plate</li>
 +
                <li>Wait for 2 days until results show up</li>
 +
              </ul>
 +
            </li>
 +
          </ul>
 +
 +
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        <h1>
 +
          Experiment
 +
        </h1>
 +
        <ul style="list-style-type: decimal;">
 +
          <li>
 +
            Confirming that indeed these mealworms digest plastic and recording how much they eat in a day
 +
          </li>
 +
          <li>
 +
            Chemical Transformation of Colon Bacteria by the Addition of PET-rase enzyme DNA (incomplete)
 +
          </li>
 +
        </ul>
 +
    </div>
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Revision as of 08:32, 18 September 2016

Experiments

Main Subtitle

Protocol

  • Mealworm Dissection
  • LB Agar Medium Synthesis

    Materials Needed:

    • Water(purified water)
    • LB broth (salt, yeast, 카세린), since we’re making a 500ml solution, we’ll need 12.5g
      • 25g/liter
    • AGAR (not easily disintegrated by bacteria, structure preserved)
      • Has to be 1.5% of the solution
      • Since we’re making a 500ml solution, we’ll need 7.5g
    • Autoclave
      • High pressure, high temp sterilization
      • More than 20 minutes

    Procedure:

    • Put water up to 500ml in a graduated cylinder pour it in a large flask
    • Measure LB broth carefully to match 12.5g inside the a plastic temporary container
    • Close the lid to the LB broth, wash the spoon used to transfer it
    • Fold the plastic container in half and pour the LB broth into the flask
    • In the same manner, put AGAR(7.5g) inside the flask
    • Don’t mix he flask, and seal the top of the flask with a tin foil
    • Put it in the autoclave, pour some hot water and close the lid wait for an hour and half
    • When storing the AGAR medium, make sure to preserve it in 60 degree cel.
    • Go to the clean bench, or if it’s unavailable you kindle the alcohol lamp(which causes convection and so in a sense forms an umbrella that protects the plate from contamination from the air)
    • Pour the AGAR solution onto eight plates, making sure your hand or your other body part is not hovering over the plate (you always have to keep in mind that SOMETHING’s falling from the air.
    • Close the lids of the all eight plates and wait till they harden.
  • Chemical Transformation
    • Basic Knowledge:
      • Competent cell
      • e-coli’s name : DH5-alpha
      • DNA to be Added : PUC19

    Procedure:

    • Prepare two containers
      • only the DH5-alpha
      • DH5 alpha + PUC19 (experimental group)
    • Put both containers in ice for a minute
    • Raise the temperature abruptly to 42 degrees Cel for a minute
    • Put them into ice for another 2 minutes
    • At the Clean Bench, put LB Medium equally to each container
    • 37°C 180 RPM (at least 30-min)
    • In the centrifuge, 30 seconds at highest speed
    • With a Pipet eliminate remaining precipitate
    • In the LB Medium plate, put appropriate number of glass beads
    • After mixing the substances in the two containers with the pipet,
    • Put the two substances into each separate LB plates, put the lid on top and shake
    • Seal the plate
    • Wait for 2 days until results show up

Experiment

  • Confirming that indeed these mealworms digest plastic and recording how much they eat in a day
  • Chemical Transformation of Colon Bacteria by the Addition of PET-rase enzyme DNA (incomplete)