Difference between revisions of "Team:Calgary/Demonstrate"

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<h1>Demonstration of our work</h1>
 
<h1>Demonstration of our work</h1>
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<h2>Removal of chlorophyll from green seed canola oil</h2>
 
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<p>To show we produced 6GIX in significant amounts, we ran SDS-PAGE gel which showed the right bands. We purified the protein using 6X His affinity chromatography and showed bands of the correct size which also confirms successful production. We also did a Western Blot. We also measured the absorption of the protein at 200nm to quantify that we have THICC amounts.</p>
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<p>Using an IPTG-inducible system, our team was able to successfully produce and purify 6GIX in <i>E.coli</i> BL21(DE3). An SDS-PAGE gel with whole cell lysate and purified elutions of 6GIX without a signal peptide showed strong bands of the correct size in the whole cell lysate and elution 2. No bands were seen in the corresponding empty vector control lanes. </p>
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<img style="display: block; margin-left: auto; margin-right: auto;" width= 100% src="https://static.igem.org/mediawiki/2019/4/48/T--Calgary--6GIXgel.jpeg"></img>
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<p style="text-align: center ;">Figure 5: SDS-PAGE gel showing whole cell lysate and elutions after purification of no signal peptide 6GIX and empty vector control. Arrow denotes correct band length for 6GIX protein.</p>
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<p>In order to ensure that the bands we see are of the correct protein, we ran a western blot using ____what antibodies and shit did we use____. The western blot also showed a band of the correct size
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<h1>Emulsification and Extraction</h1>
 
<h1>Emulsification and Extraction</h1>

Revision as of 20:02, 20 October 2019

sub-title

Demonstrate

Demonstration of our work

Removal of chlorophyll from green seed canola oil

Our team’s goal was to create a new method for chlorophyll removal from green seed canola oil. After months of hard work and experimentation, our team was able to demonstrate the following:

  1. Our water soluble chlorophyll-binding protein, 6GIX, can be produced and purified
  2. 6GIX can be emulsified and can be used to remove chlorophyll from canola oil

6GIX Production

Using an IPTG-inducible system, our team was able to successfully produce and purify 6GIX in E.coli BL21(DE3). An SDS-PAGE gel with whole cell lysate and purified elutions of 6GIX without a signal peptide showed strong bands of the correct size in the whole cell lysate and elution 2. No bands were seen in the corresponding empty vector control lanes.

Figure 5: SDS-PAGE gel showing whole cell lysate and elutions after purification of no signal peptide 6GIX and empty vector control. Arrow denotes correct band length for 6GIX protein.

In order to ensure that the bands we see are of the correct protein, we ran a western blot using ____what antibodies and shit did we use____. The western blot also showed a band of the correct size

Emulsification and Extraction

We proved that our emulsification system successfully reduces the amount of chlorophyll in canola oil. with this THICC experiment