Difference between revisions of "Team:Hong Kong-CUHK/Results"

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<h1>Result </h1>
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      <a class="navbar-brand" href="#">Results</a>
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          <li><a href="#section1">Toehold Switch</a></li>
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          <li><a href="#section2">Interlab</a></li>
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 Influenza switches
  
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  <h1>Toehold Switch</h1>
 
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  <h1>Interlab</h1>
 
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  <p>Please refer to the Interlab part.</p>
 
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  <h1>Characterization</h1>
 
  
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We found that amajLime was described as “chromoprotein” in the main page. We later found that amajLime is a fluorescent protein instead. Therefore we scanned the emission and excitation spectrum of it and documented it in registry:
 
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<img src="https://static.igem.org/mediawiki/parts/1/15/Ex-Em._amajlime.png" width="50%" height="auto">
 
  
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We then measured the fluorescent signal of mRFP and amajLime in buffer with different pH:
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 Toehold switch cloning tool
  
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<img src="
 
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https://static.igem.org/mediawiki/parts/5/56/Mrfp.PNG" width="50%" height="auto">
 
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<img src="https://static.igem.org/mediawiki/parts/4/44/Amaj.PNG" width="50%" height="auto">
 
  
  
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Purified mRFP and amajLime was diluted to 10µg/100µl (total 200µl), in triplicates, into different buffers which pH ranges from pH2 to pH12. Volume of mRFP: pH buffer=1:1.8, amajLime : pH buffer =1:4.3. The lines represent the trend of the median fluorescence for the measured pH values. mRFP and amajLime were excited by 584nm and 470nm respectively and emission was measured at 607nm and 515nm respectively. To allow repeatable experiment, we converted the relative fluorescent intensity to an absolute fluorophore concentration by plotting standard curve of the fluorophore using the interlab protocol. We used Rhodamine to measure mRFP; Fluorescein to measure amajLime. The results show that the optimum pH of both chromoprotein ranges from pH6 to pH8. Their fluorescent signal thus will probably not be interfered by the pH of our proposed sample: saliva or NP swab.
 
  
  
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 Improving existing biobricks and project: Cancer switches
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  <h1>Program</h1>
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<h3>Characterization of chromoprotein</h3>
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We found that amajLime was described as “chromoprotein” in the main page. We later found that amajLime is a fluorescent protein instead. Therefore we scanned the emission and excitation spectrum of it and documented it in registry:
 +
<br>
 +
<img src="https://static.igem.org/mediawiki/parts/1/15/Ex-Em._amajlime.png" width="50%" height="auto">
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<br>
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We then measured the fluorescent signal of mRFP and amajLime in buffer with different pH:
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<br>
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<img src="
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https://static.igem.org/mediawiki/parts/5/56/Mrfp.PNG" width="50%" height="auto">
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<br>
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<img src="https://static.igem.org/mediawiki/parts/4/44/Amaj.PNG" width="55%" height="auto">
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<br>
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Purified mRFP and amajLime was diluted to 10µg/100µl (total 200µl), in triplicates, into different buffers which pH ranges from pH2 to pH12. Volume of mRFP: pH buffer=1:1.8, amajLime : pH buffer =1:4.3. The lines represent the trend of the median fluorescence for the measured pH values. mRFP and amajLime were excited by 584nm and 470nm respectively and emission was measured at 607nm and 515nm respectively. To allow repeatable experiment, we converted the relative fluorescent intensity to an absolute fluorophore concentration by plotting standard curve of the fluorophore using the interlab protocol. We used Rhodamine to measure mRFP; Fluorescein to measure amajLime. The results show that the optimum pH of both chromoprotein ranges from pH6 to pH8. Their fluorescent signal thus will probably not be interfered by the pH of our proposed sample: saliva or NP swab.
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Revision as of 12:39, 22 October 2017





Result

 Influenza switches  Toehold switch cloning tool  Improving existing biobricks and project: Cancer switches

Characterization of chromoprotein

We found that amajLime was described as “chromoprotein” in the main page. We later found that amajLime is a fluorescent protein instead. Therefore we scanned the emission and excitation spectrum of it and documented it in registry:

We then measured the fluorescent signal of mRFP and amajLime in buffer with different pH:


Purified mRFP and amajLime was diluted to 10µg/100µl (total 200µl), in triplicates, into different buffers which pH ranges from pH2 to pH12. Volume of mRFP: pH buffer=1:1.8, amajLime : pH buffer =1:4.3. The lines represent the trend of the median fluorescence for the measured pH values. mRFP and amajLime were excited by 584nm and 470nm respectively and emission was measured at 607nm and 515nm respectively. To allow repeatable experiment, we converted the relative fluorescent intensity to an absolute fluorophore concentration by plotting standard curve of the fluorophore using the interlab protocol. We used Rhodamine to measure mRFP; Fluorescein to measure amajLime. The results show that the optimum pH of both chromoprotein ranges from pH6 to pH8. Their fluorescent signal thus will probably not be interfered by the pH of our proposed sample: saliva or NP swab.
     

Email: igemcuhk1617@gmail.com