Difference between revisions of "Team:Lanzhou/Demonstrate"

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$('.page__header').css('background-image', 'url(https://static.igem.org/mediawiki/2017/2/24/Lanzhou_wiki_Header_background.jpg)');
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$('.page__header h1').text('Results');
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<div class="column full_size judges-will-not-evaluate">
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<article id="article_1" class="page__article">
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<h3>★  ALERT! </h3>
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<h1 class="mdc-typography--headline">DsRNA Extraction</h1>
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<p>This page is used by the judges to evaluate your team for the <a href="https://2017.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2017.igem.org/Judging/Awards"> award listed above</a>. </p>
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<p class="mdc-typography--body2">
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<p> Delete this box in order to be evaluated for this medal criterion and/or award. See more information at <a href="https://2017.igem.org/Judging/Pages_for_Awards"> Instructions for Pages for awards</a>.</p>
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We extracted dsTrxz/hpTrxz produced by M-JM109-LacY containing BBa_K2377004 (Fig.1)/BBa_K2377005 (Fig.2). M-JM109-LacY and M-JM109-LacY containing pDP (Fig.3) are control groups.
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<img src="https://static.igem.org/mediawiki/2017/4/45/Lanzhou_wiki_Results_figure1.png" alt="" data-action="zoom">
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<figcaption>Fig.1 BBa_K2377004: DPTrxz</figcaption>
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<div class="column full_size">
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</figure>
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<h1>Demonstrate</h1>
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<figure>
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<h3>Gold Medal Criterion #4</h3>
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<img src="https://static.igem.org/mediawiki/2017/6/60/Lanzhou_wiki_Results_figure2.png" alt="" data-action="zoom">
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<figcaption>Fig.2 BBa_K2377005: TrxzL</figcaption>
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<p>
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</figure>
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Teams that can show their system working under real world conditions are usually good at impressing the judges in iGEM. To achieve gold medal criterion #4, convince the judges that your project works. There are many ways in which your project working could be demonstrated, so there is more than one way to meet this requirement. This gold medal criterion was introduced in 2016, so check our what 2016 teams did to achieve a their gold medals!
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<figure>
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</p>
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<img src="https://static.igem.org/mediawiki/2017/5/5d/Lanzhou_wiki_Results_figure3.png" alt="" data-action="zoom">
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<figcaption>Fig.3 BBa_K2377016: pDP</figcaption>
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<p>
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</figure>
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Please see the <a href="https://2017.igem.org/Judging/Medals">2017 Medals Page</a> for more information.
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<p class="mdc-typography--body2">
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Products are shown in Fig.4. DsTrxz and hpTrxz are approximately 500bp. We can see in the picture that dsTrxz and hpTrxz are successfully extracted. (Yellow box)
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<figure>
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<img src="https://static.igem.org/mediawiki/2017/2/2c/Lanzhou_wiki_Results_figure4.jpg" alt="" data-action="zoom">
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<figcaption>
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Fig.4 Gel electrophoresis picture of dsRNA extraction products. (1) BBa_K2377004: DPTrxz, mini-preparation. (2) BBa_K2377005: TrxzL, mini-preparation. (3) BBa_K2377004: DPTrxz, mass isolation. (4) BBa_K2377005: TrxzL, mass isolation. (5) BBa_K2377016: pDP mini-preparation. (6) M-JM109-LacY, mini-preparation. We can see in the picture that dsTrxz and hpTrxz are successfully extracted. (yellow box)
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<h4> What should we do for our demonstration?</h4>
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<h1 class="mdc-typography--headline">Plant Treatment</h1>
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<p class="mdc-typography--body2">
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<h5> Standard teams </h5>
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To verify the function of dsTrxz, we did the plant treatment experiment.
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</p>
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<p>  
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<p class="mdc-typography--body2">
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If you have built a proof of concept system, you can demonstrate it working under real world conditions. If you have built a biological device that is intended to be a sensor, can you show it detecting whatever it is intended to sense. If it is intended to work in the field, you can show how this might work using a simulated version in the lab, or a simulation of your device in the field.<strong> Please note biological materials must not be taken out of the lab</strong>.
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Three groups of Arabidopsis are shown below:
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<thead>
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<br>
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<th>group/treatment</th>
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<h5> Special track teams </h5>
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<th>Days in MS media</th>
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<th>Days in MS + dsTrxz</th>
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<p>
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</tr>
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Special track teams can achieve this medal criterion by bringing their work to the Jamboree and showcasing it in the track event. Art & Design, Measurement, Hardware and Software tracks will all have showcase events at the Giant Jamboree.<strong> Please note biological materials must not be taken out of the lab</strong>.
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<tbody>
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<th>group 1</th>
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<td>7</td>
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<td>5</td>
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</tr>
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<tr>
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<th>group 2</th>
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<td>14</td>
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<td>21</td>
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</tr>
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<tr>
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<th>group 3</th>
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<td>22</td>
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<td>30</td>
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</tbody>
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</table>
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<figcaption>Data of group 2 is perfect but lost. Here we show the data of group 1 and group 3.</figcaption>
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</figure>
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<h2 class="mdc-typography--title">Group 1</h2>
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<p class="mdc-typography--body2">
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We seperated group 1 into six subgroups. Each group is treated with different concentration of dsTrxz:
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</p>
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<figure>
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<table>
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<thead>
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<tr>
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<th>Subgroup</th>
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<th>Concentration (ng/L)</th>
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</tr>
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</thead>
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<tbody>
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<tr>
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<th>1</th>
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<td>0</td>
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</tr>
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<tr>
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<th>2</th>
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<td>0.1</td>
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</tr>
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<tr>
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<th>3</th>
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<td>1</td>
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<th>4</th>
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<td>10</td>
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<tr>
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<th>5</th>
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<td>100</td>
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</tr>
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<tr>
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<th>6</th>
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<td>1000</td>
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</tr>
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</tbody>
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</table>
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</figure>
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<p class="mdc-typography--body2">
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We did this experiment to optimize concentration of dsTrxz treatment. As we can see in figure 5, LWRs with dsTrxz treatment are higher than those without dsTrxz treatment. LWRs are relative stable with consentration higher than 1ng/L.
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</p>
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<figure>
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<img src="https://static.igem.org/mediawiki/2017/7/78/Lanzhou_wiki_Results_figure5.png" alt="" data-action="zoom">
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<figcaption>
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Fig.5 Average Leaf Whiting Rate (LWR) of Arabidopsis 5 days after growing in media with different concentration of dsTrxz.
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</figcaption>
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</figure>
  
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<h2 class="mdc-typography--title">Group 2</h2>
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<p class="mdc-typography--body2">
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The picture (Fig. 6) of Control group and dsTrxz (+) group shows significant differeces caused by dsTrxz treatment (10ng/L).
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</p>
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<figure>
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<img src="https://static.igem.org/mediawiki/2017/thumb/1/10/Lanzhou_wiki_Results_figure6.png/800px-Lanzhou_wiki_Results_figure6.png" alt="" data-action="zoom">
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<figcaption>
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Fig.6 Comparison of control group and treatment group. 2a. Treatment group shows significant poorer growth vigor than control group. 2b. Leaves turn yellow one month after dsTrxz treatment.
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</figcaption>
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</figure>
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<p class="mdc-typography--body2">
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We measured number of leaves (Fig.7a), average length of longest leaves (Fig. 7b) and average LWR (Fig. 7c) of contol group and dsTrxz group. Results show significant differences after dsTrxz treatment.
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</p>
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<figure style="max-width=90%;">
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<img src="https://static.igem.org/mediawiki/2017/4/4c/Lanzhou_wiki_Results_figure7.png" alt="" data-action="zoom">
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<figcaption>
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Fig.7 Arabidopsis 1 month after growing on media with 10ng/L dsTrxz. (a) Number of leaves shows changes of growth vigor after dsTrxz treatment. (b) Average length of longest leaves shows changes of growth vigor agter dsTrxztreatment. (c) Average LWR shows significant whiting phenotype after dsTrxz treatment.
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</figcaption>
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</figure>
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</article>
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{{Lanzhou/Footer}}

Latest revision as of 02:54, 2 November 2017

Lanzhou

Lanzhou2017

DsRNA Extraction

We extracted dsTrxz/hpTrxz produced by M-JM109-LacY containing BBa_K2377004 (Fig.1)/BBa_K2377005 (Fig.2). M-JM109-LacY and M-JM109-LacY containing pDP (Fig.3) are control groups.

Fig.1 BBa_K2377004: DPTrxz
Fig.2 BBa_K2377005: TrxzL
Fig.3 BBa_K2377016: pDP

Products are shown in Fig.4. DsTrxz and hpTrxz are approximately 500bp. We can see in the picture that dsTrxz and hpTrxz are successfully extracted. (Yellow box)

Fig.4 Gel electrophoresis picture of dsRNA extraction products. (1) BBa_K2377004: DPTrxz, mini-preparation. (2) BBa_K2377005: TrxzL, mini-preparation. (3) BBa_K2377004: DPTrxz, mass isolation. (4) BBa_K2377005: TrxzL, mass isolation. (5) BBa_K2377016: pDP mini-preparation. (6) M-JM109-LacY, mini-preparation. We can see in the picture that dsTrxz and hpTrxz are successfully extracted. (yellow box)

Plant Treatment

To verify the function of dsTrxz, we did the plant treatment experiment.

Three groups of Arabidopsis are shown below:

group/treatment Days in MS media Days in MS + dsTrxz
group 1 7 5
group 2 14 21
group 3 22 30
Data of group 2 is perfect but lost. Here we show the data of group 1 and group 3.

Group 1

We seperated group 1 into six subgroups. Each group is treated with different concentration of dsTrxz:

Subgroup Concentration (ng/L)
1 0
2 0.1
3 1
4 10
5 100
6 1000

We did this experiment to optimize concentration of dsTrxz treatment. As we can see in figure 5, LWRs with dsTrxz treatment are higher than those without dsTrxz treatment. LWRs are relative stable with consentration higher than 1ng/L.

Fig.5 Average Leaf Whiting Rate (LWR) of Arabidopsis 5 days after growing in media with different concentration of dsTrxz.

Group 2

The picture (Fig. 6) of Control group and dsTrxz (+) group shows significant differeces caused by dsTrxz treatment (10ng/L).

Fig.6 Comparison of control group and treatment group. 2a. Treatment group shows significant poorer growth vigor than control group. 2b. Leaves turn yellow one month after dsTrxz treatment.

We measured number of leaves (Fig.7a), average length of longest leaves (Fig. 7b) and average LWR (Fig. 7c) of contol group and dsTrxz group. Results show significant differences after dsTrxz treatment.

Fig.7 Arabidopsis 1 month after growing on media with 10ng/L dsTrxz. (a) Number of leaves shows changes of growth vigor after dsTrxz treatment. (b) Average length of longest leaves shows changes of growth vigor agter dsTrxztreatment. (c) Average LWR shows significant whiting phenotype after dsTrxz treatment.