Difference between revisions of "Team:NTU-Singapore/Measurement"

 
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    <section id="a1" class="home-section-humanprac text-center bg-gray" >
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<div id="rbs" class="inner tittle" align="center" >
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<p style="font-size: 30px; color: white;">Ribosomal Binding Site</p>
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</div>
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<div class="container" id="rbs">
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        <div class="row boxed-grey-humanprac"  >
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<div class="wow bounceInUp" data-wow-delay="0.2s" >
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<div class="inner" align="center">
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<h5>Growth Curve</h5>
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<p class="subtitle explain" style=" width: 1200px; ">
 +
                                                       
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As the measurements are carried out in six batches, the growth of mutants of the same batch are similar but differed a little among batches. This implies that GFP expression does not have any significant effect on the bacteris's growth.
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<br><br>
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1AT denotes A of base pair 1 is changed to T, the same is applied for other notations of RBS mutants.
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<br><br>
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<img src="https://static.igem.org/mediawiki/2015/1/1f/123_gc.png" width="400" height="372"><img src="https://static.igem.org/mediawiki/2015/f/f5/789gc.png"width="400" height="372">
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<img src="https://static.igem.org/mediawiki/2015/9/9e/456.png"width="390" height="372"> <br><img src="https://static.igem.org/mediawiki/2015/1/1b/10%2C11%2C12gc.png"width="400" height="372">
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<img src="https://static.igem.org/mediawiki/2015/9/93/6gt.png"width="400" height="372"><img src="https://static.igem.org/mediawiki/2015/2/2a/108GC.png"width="390" height="372">
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</div>
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        </div>
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<br>
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</div>
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</div>
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</section>
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<!--Activity2-->
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<section id="a2" class="home-section-humanprac text-center">
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<div class="container">
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<div class="row boxed-grey-humanprac"  >
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<div class="wow bounceInUp" data-wow-delay="0.2s">
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 +
<div class="inner" align="center">
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<h5>GFP Readings</h5>
 +
<p class="subtitle explain">
 +
The GFP fluorescence readings of mutants shows interesting results. Although the growth curve is similar among mutants, fluorescence intensities varied among the mutants. In summary, it was found that substitution mutations occurring to the AGGAG sequence within BBa_R0034, AAAGAGGAGAAA, showed a decreased GFP expression while others showed increased GFP output. Especially for mutations to base pair 7, GFP expression is near total-depression.
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</p>
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<img src="https://static.igem.org/mediawiki/2015/8/81/123.png"><img src="https://static.igem.org/mediawiki/2015/4/4f/Gfp456.png"><img src="https://static.igem.org/mediawiki/2015/7/78/Gfp6gt.png"><img src="https://static.igem.org/mediawiki/2015/9/93/Gfp789.png"><img src="https://static.igem.org/mediawiki/2015/8/82/Gfp10%2C8.png"><img src="https://static.igem.org/mediawiki/2015/2/22/Gfp101112.png"><br><br>
 +
<p class="subtitle explain">
 +
After normalising the GFP fluorescence readings with the OD<sub>600</sub> at T=8, the ratio of the normalised fluorescence of the mutants to wild type RBS is computed and plotted as shown.
 +
<br><br>
 +
<img src="https://static.igem.org/mediawiki/2015/9/90/Sumary.png" style="margin-left: 350px;">
 +
                                                </p>
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</div>
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</div>
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</div>
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<br>
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</div>
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</div>
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</section>
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<section id="a2" class="home-section-humanprac text-center">
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<div class="container">
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 +
 +
<div class="row boxed-grey-humanprac"  >
 +
 +
<div class="wow bounceInUp" data-wow-delay="0.2s">
 +
 +
<div class="inner" align="center">
 +
<h5>Spotting</h5>
 +
<p class="subtitle explain">
 +
We also spotted the mutants on an LB agar plate to have a qualitative view of the GFP brightness. The culture is diluted to OD<sub>600</sub> = 0.4 then 3uL of the culture is spotted on to the agar. Brightness of these spots parallels the results in the above graph. For example, 7GA, 8AG and 9GA is the brightest among the mutations on their respective base pairs while 12AC and 11AC are the darkest. <div align="centre"><img class="" src="https://static.igem.org/mediawiki/2015/3/3a/Rbs-spots.jpeg" width="821.6px" height="550px"></div>
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<br><br>
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                                                </p>
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</div>
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</div>
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    <section id="a1" class="home-section-humanprac text-center bg-gray" >
 
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<div id="rbs" class="inner tittle" align="center" >
 
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<p style="font-size: 30px; color: white;">Ribosomal Binding Site</p>
 
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</div>
 
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<div class="container" id="rbs">
 
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        <div class="row boxed-grey-humanprac"  >
 
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<div class="wow bounceInUp" data-wow-delay="0.2s" >
 
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<div class="inner" align="center">
 
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<h5>Growth Curve</h5>
 
−
<p class="subtitle explain" style=" width: 1200px; ">
 
−
                                                       
 
−
As the measurements are carried out in six batches, the growth of mutants of the same batch are similar but differed a little among batches. This implies that GFP expression does not have any significant effect on the bacteris's growth.
 
−
 
−
<br><br>
 
−
 
−
1AT denotes A of base pair 1 is changed to T, the same is applied for other notations of RBS mutants.
 
−
 
−
<br><br>
 
−
 
−
<img src="https://static.igem.org/mediawiki/2015/1/1f/123_gc.png" width="400" height="372"><img src="https://static.igem.org/mediawiki/2015/f/f5/789gc.png"width="400" height="372">
 
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<img src="https://static.igem.org/mediawiki/2015/9/9e/456.png"width="390" height="372"> <br><img src="https://static.igem.org/mediawiki/2015/1/1b/10%2C11%2C12gc.png"width="400" height="372">
 
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<img src="https://static.igem.org/mediawiki/2015/9/93/6gt.png"width="400" height="372"><img src="https://static.igem.org/mediawiki/2015/2/2a/108GC.png"width="390" height="372">
 
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<div class="inner" align="center">
 
−
<h5>GFP Readings</h5>
 
−
<p class="subtitle explain">
 
−
The GFP fluorescence readings of mutants shows interesting results. Although the growth curve is similar among mutants, fluorescence intensities varied among the mutants. In summary, it was found that substitution mutations occurring to the AGGAG sequence within BBa_R0034, AAAGAGGAGAAA, showed a decreased GFP expression while others showed increased GFP output. Especially for mutations to base pair 7, GFP expression is near total-depression.
 
−
</p>
 
−
<img src="https://static.igem.org/mediawiki/2015/8/81/123.png"><img src="https://static.igem.org/mediawiki/2015/4/4f/Gfp456.png"><img src="https://static.igem.org/mediawiki/2015/7/78/Gfp6gt.png"><img src="https://static.igem.org/mediawiki/2015/9/93/Gfp789.png"><img src="https://static.igem.org/mediawiki/2015/8/82/Gfp10%2C8.png"><img src="https://static.igem.org/mediawiki/2015/2/22/Gfp101112.png"><br><br>
 
−
<p class="subtitle explain">
 
−
After normalising the GFP fluorescence readings with the OD<sub>600</sub> at T=8, the ratio of the normalised fluorescence of the mutants to wild type RBS is computed and plotted as shown.
 
−
<br><br>
 
−
<img src="https://static.igem.org/mediawiki/2015/9/90/Sumary.png" style="margin-left: 350px;">
 
−
                                                </p>
 
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<div class="row boxed-grey-humanprac"  >
 
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<div class="wow bounceInUp" data-wow-delay="0.2s">
 
−
 
−
<div class="inner" align="center">
 
−
<h5>Spotting</h5>
 
−
<p class="subtitle explain">
 
−
We also spotted the mutants on an LB agar plate to have a qualitative view of the GFP brightness. The culture is diluted to OD<sub>600</sub> = 0.4 then 3uL of the culture is spotted on to the agar. Brightness of these spots parallels the results in the above graph. For example, 7GA, 8AG and 9GA is the brightest among the mutations on their respective base pairs while 12AC and 11AC are the darkest. <div align="centre"><img class="" src="https://static.igem.org/mediawiki/2015/3/3a/Rbs-spots.jpeg" width="821.6px" height="550px"></div>
 
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Latest revision as of 17:27, 18 September 2015

NTU SG iGEM 2015




Measurement

Ribosomal Binding Site

Growth Curve

As the measurements are carried out in six batches, the growth of mutants of the same batch are similar but differed a little among batches. This implies that GFP expression does not have any significant effect on the bacteris's growth.

1AT denotes A of base pair 1 is changed to T, the same is applied for other notations of RBS mutants.



GFP Readings

The GFP fluorescence readings of mutants shows interesting results. Although the growth curve is similar among mutants, fluorescence intensities varied among the mutants. In summary, it was found that substitution mutations occurring to the AGGAG sequence within BBa_R0034, AAAGAGGAGAAA, showed a decreased GFP expression while others showed increased GFP output. Especially for mutations to base pair 7, GFP expression is near total-depression.



After normalising the GFP fluorescence readings with the OD600 at T=8, the ratio of the normalised fluorescence of the mutants to wild type RBS is computed and plotted as shown.


Spotting

We also spotted the mutants on an LB agar plate to have a qualitative view of the GFP brightness. The culture is diluted to OD600 = 0.4 then 3uL of the culture is spotted on to the agar. Brightness of these spots parallels the results in the above graph. For example, 7GA, 8AG and 9GA is the brightest among the mutations on their respective base pairs while 12AC and 11AC are the darkest.