Difference between revisions of "Team:ASTWS-China/Improve"

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        <h1>Improve</h1>
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        <p style="text-align: center"><b><i>Part: <a href="http://parts.igem.org/Part:BBa_K2826008">BBa_K2826008</a></i></b></p>
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        <h2>Description &amp; Design</h2>
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        <p>This part is an improvement of part <a href="http://parts.igem.org/Part:BBa_K2088015">BBa_K2088015</a>, which consisted by the same copper ion detecting promoter and GFP as reporter. Comparing with BBa_K2088015, our improvement is using RFP to replace GFP as reporter. The composition of part BBa_K2826008 is shown in Figure 1.</p>
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        <img src="https://static.igem.org/mediawiki/2018/6/6f/T--ASTWS-China--improve1.jpg">
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        <p id="annotation">Figure 1: Composition of part BBa_K2826008</p>
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        <h2>Experimental method</h2>
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        <p>2.5 ml of LB solution was added with 2.5 ml of sterilized ultrapure water, and BBa_K2826013 broth was added to make a final concentration of 0.02 OD, cultured at 37 °C on a shaker for 10 hours. 100 μL of bacterial solution was sampled every 2 hours to detect red fluorescence intensity. When measuring wastewater, replace 2.5 ml of ultrapure water with 2.5 ml of waste water.</p>
  
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        <h2>Results and Discussion</h2>
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<h3>★  ALERT! </h3>
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        <p>As shown in Figure 2, with the extension of culture time, the fluorescence intensity of eRFP gradually increased which indicates that the expression level of eRFP in the medium gradually increased. In other words, the copper detection function of this working system was working normally. The concentration of copper ions in the waste water is close to 3ml/L. The detection results are similar to measurement result of ICP. Besides, as shown in Figure 3, as the copper concentration increased, the color turned redder. It is proved that the copper detection function of this working system was working as we expected.</p>
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<p>This page is used by the judges to evaluate your team for the <a href="https://2018.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2018.igem.org/Judging/Awards"> award listed below</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://2018.igem.org/Judging/Pages_for_Awards"> Instructions for Pages for awards</a>.</p>
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        <p id="annotation">Figure 2: Curve of fluorescence intensity and culture time under different copper concentrations</p>
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        <p id="annotation">Figure 3: E.coli DH5α after culturing for 10 hours under different copper concentrations.</p>
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        <h2>Conclusion</h2>
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<h1>Improve</h1>
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        <p>This design modification is based on three reasons and has significant advantage:</p>
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<p>For teams seeking to improve upon a previous part or project, you should document all of your work on this page. Please remember to include all part measurement and characterization data on the part page on the Registry. Please include a link to your improved part on this page.</p>
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        <p>1. Comparing with GFP, RFP has a visible color which can be observed by naked eyes without UV excitation, therefore, in same condition, our new part can be used for qualitatively easily. Moreover, since it is not necessary to have a UV light source for qualitatively detecting, the prototype device could be smaller and cheaper.</p>
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        <p>2. RFP fluoresces red-orange when excited, therefore, in case of quantitative use, it will be same as GFP.</p>
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<h3>Gold Medal Criterion #2</h3>
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        <p>3. Usually, red color means “dangerous” and green color means “safe”. In the case of heavy metal pollution detection, the detecting result means more “dangerous” and “alert”, thus, using red color instead of green is in keeping with common sense.</p>
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<p><b>Standard Tracks:</b> Create a new part that has a functional improvement upon an existing BioBrick part. The sequences of the new and existing parts must be different. You must perform experiments with both parts to demonstrate this improvement. Document the experimental characterization on the Part's Main Page on the Registry for both the existing and new parts. Both the new and existing Main Page of each Part’s Registry entry must reference each other. Submit a sample of the new part to the Registry.
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The existing part must NOT be from your 2018 part number range and must be different from the part documented in bronze #4.
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<b>Special Tracks:</b> Improve the function of an existing iGEM project (that your current team did not originally create) and display your achievement on your wiki.</p>
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                        <p class="alignleft">Copyright &#169; 2018 Team:ASTWS-China All Rights Reserved.</p>
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Latest revision as of 20:02, 17 October 2018

Improve

Part: BBa_K2826008

Description & Design

This part is an improvement of part BBa_K2088015, which consisted by the same copper ion detecting promoter and GFP as reporter. Comparing with BBa_K2088015, our improvement is using RFP to replace GFP as reporter. The composition of part BBa_K2826008 is shown in Figure 1.

Figure 1: Composition of part BBa_K2826008

Experimental method

2.5 ml of LB solution was added with 2.5 ml of sterilized ultrapure water, and BBa_K2826013 broth was added to make a final concentration of 0.02 OD, cultured at 37 °C on a shaker for 10 hours. 100 μL of bacterial solution was sampled every 2 hours to detect red fluorescence intensity. When measuring wastewater, replace 2.5 ml of ultrapure water with 2.5 ml of waste water.

Results and Discussion

As shown in Figure 2, with the extension of culture time, the fluorescence intensity of eRFP gradually increased which indicates that the expression level of eRFP in the medium gradually increased. In other words, the copper detection function of this working system was working normally. The concentration of copper ions in the waste water is close to 3ml/L. The detection results are similar to measurement result of ICP. Besides, as shown in Figure 3, as the copper concentration increased, the color turned redder. It is proved that the copper detection function of this working system was working as we expected.

Figure 2: Curve of fluorescence intensity and culture time under different copper concentrations

Figure 3: E.coli DH5α after culturing for 10 hours under different copper concentrations.

Conclusion

This design modification is based on three reasons and has significant advantage:

1. Comparing with GFP, RFP has a visible color which can be observed by naked eyes without UV excitation, therefore, in same condition, our new part can be used for qualitatively easily. Moreover, since it is not necessary to have a UV light source for qualitatively detecting, the prototype device could be smaller and cheaper.

2. RFP fluoresces red-orange when excited, therefore, in case of quantitative use, it will be same as GFP.

3. Usually, red color means “dangerous” and green color means “safe”. In the case of heavy metal pollution detection, the detecting result means more “dangerous” and “alert”, thus, using red color instead of green is in keeping with common sense.

Copyright © 2018 Team:ASTWS-China All Rights Reserved.