Difference between revisions of "Team:ITB Indonesia/Contribution"

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{{ITB_Indonesia}}
 
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<h3>★  ALERT! </h3>
 
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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> 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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<div class="column full_size" style="background: #1c2922; margin-top: -40px; padding: 30px;">
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<h1 class="ITB_h1" style="margin-bottom: 50px;">Contribution</h1>
  
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<h1>Contribution</h1>
 
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<h3>Bronze Medal Criterion #4</h3>
 
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<p><b>Standard Tracks:</b> Participate in the Interlab Measurement Study (to be documented on your InterLab page) and/or improve the characterization of an existing BioBrick Part or Device and enter this information on that part's Main Page in the Registry. The part that you are characterizing must NOT be from a 2017 part number range. Teams who are working on improving the characterization of an existing part should document their experimental design here, along with an explanation for why they chose that part to improve. Data can also be shown here, but it MUST also be documented on the part's Main Page in the Registry.
 
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<br><br>
 
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<b>Special Tracks:</b> Document at least one new substantial contribution to the iGEM community that showcases a project related to BioBricks. This contribution should be central to your project and equivalent in difficulty to making and submitting a BioBrick part.
 
  
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            <div style="background: #e8e6d1; padding: 30px;">
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<h1 class="ITB_h1" style="margin-bottom: 50px; color: #1c2922">New characterization to RFP Coding Device</h1>
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<figure>
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                                                          <img src="
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                    https://static.igem.org/mediawiki/parts/5/53/Barenganstrain.JPG" style="height: 75%; width: 75%;"/>
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                                    <figcaption><i><b>Figure 1:</b> There is no significant difference of mRFP expression between three strains (Bl21, DH5alpha, Top10)</i></figcaption>
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                                                        </figure>
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<p><i><strong>Background</strong></i><br><br>
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In <a href="https://2017.igem.org/Team:ITB_Indonesia" target="_blank">Team ITB_Indonesia 2017</a>
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characterization, we found in normal growth/incubation condition (37 <sup>o</sup>C, LB agar) that
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<a href="http://parts.igem.org/Part:BBa_J04450">BBa_J04450</a>
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-transformed <i>Escherichia coli</i> BL21 colony appear to need longer incubation time (>18
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hours) until it clearly shows red color under natural light.<br><br>
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We then investigate whether this phenomenon is influenced by the strain, and we try if there are
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<i>lac</i> repressor in the system that can be released by inducing the culture with IPTG, hence
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increasing the expression of mRFP.<br><br>
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<i><strong>Experimental Design</strong></i><br><br>
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We used three different strains of transformed <i>E. coli</i> (BL21, DH5alpha, and Top10) for this
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study. They were incubated in LB broth, 37 <sup>o</sup>C, and sampled every 4 hours for 2 days to
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determine the red color absorbance at 588 nm. The amount of IPTG added for respective treatment is 500
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µM.<br><br></p>
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<p><i><strong>Result and Findings</strong></i><br>
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  <li>There are no significant differences of mRFP expression in different strains of <i>E. coli</i>
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(BL21, DH5alpha, Top10)</li>
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  <li>There are no significant effects of mRFP increased expression after IPTG induction.</li>
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  <li>The red color absorbance under 588 nm wavelength is recorded around 2.5-3 OD units.</li>
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  <li>The broth become red in color under natural light around 16-20 hours of incubation time.</li>
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</ul>
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<br>
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<figure><img src="
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                    https://static.igem.org/mediawiki/2017/0/06/Foto_mRFP1.jpg" style="height: 45%; width: 45%;"/>
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                                          <figcaption><i><b>Figure 2:</b> ~18 hours old mRFP-transformed E. coli strains (Right: control)</i></figcaption></figure>
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<figure><img src="
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                    https://static.igem.org/mediawiki/2017/2/2c/Foto_mRFP2.jpg" style="height: 45%; width: 45%;"/>
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                                          <figcaption><i><b>Figure 3:</b> ~30 hours old mRFP-transformed E. coli strains (Left: control). Notice the color turn to bright purple.</i></figcaption></figure>
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</p>
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<figure><img src="
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                    https://static.igem.org/mediawiki/parts/7/72/Top10woIPTG.JPG" style="height: 75%; width: 75%;"/>
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                                          <figcaption><i><b>Figure 4:</b> Red color dynamics of transformed E. coli Top10 compared to control</i></figcaption></figure>
  
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<figure><img src="
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                    https://static.igem.org/mediawiki/parts/b/bd/DH5woIPTG.jpeg" style="height: 75%; width: 75%;"/>
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                                          <figcaption><i><b>Figure 5:</b> Red color dynamics of transformed E. coli DH5alpha compared to control</i></figcaption></figure>
  
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<figure><img src="
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                    https://static.igem.org/mediawiki/parts/b/b7/BL21wolIPTG.jpeg" style="height: 75%; width: 75%;"/>
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                                          <figcaption><i><b>Figure 6:</b> Red color dynamics of transformed E. coli BL21 compared to control</i></figcaption></figure>
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Latest revision as of 16:50, 1 November 2017

Team Attributions
Collaborations Overview Design Notebook Contribution Results Improve
Basic Parts Composite Part Part Collection
Silver HP Integrated and Gold Public Engagement


Contribution

New characterization to RFP Coding Device

Figure 1: There is no significant difference of mRFP expression between three strains (Bl21, DH5alpha, Top10)

Background

In Team ITB_Indonesia 2017 characterization, we found in normal growth/incubation condition (37 oC, LB agar) that BBa_J04450 -transformed Escherichia coli BL21 colony appear to need longer incubation time (>18 hours) until it clearly shows red color under natural light.

We then investigate whether this phenomenon is influenced by the strain, and we try if there are lac repressor in the system that can be released by inducing the culture with IPTG, hence increasing the expression of mRFP.

Experimental Design

We used three different strains of transformed E. coli (BL21, DH5alpha, and Top10) for this study. They were incubated in LB broth, 37 oC, and sampled every 4 hours for 2 days to determine the red color absorbance at 588 nm. The amount of IPTG added for respective treatment is 500 µM.

Result and Findings

  • There are no significant differences of mRFP expression in different strains of E. coli (BL21, DH5alpha, Top10)
  • There are no significant effects of mRFP increased expression after IPTG induction.
  • The red color absorbance under 588 nm wavelength is recorded around 2.5-3 OD units.
  • The broth become red in color under natural light around 16-20 hours of incubation time.

Figure 2: ~18 hours old mRFP-transformed E. coli strains (Right: control)
Figure 3: ~30 hours old mRFP-transformed E. coli strains (Left: control). Notice the color turn to bright purple.
Figure 4: Red color dynamics of transformed E. coli Top10 compared to control
Figure 5: Red color dynamics of transformed E. coli DH5alpha compared to control
Figure 6: Red color dynamics of transformed E. coli BL21 compared to control