Difference between revisions of "Team:Waterloo/Results"

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<h1>Results </h1>  
 
<h1>Results </h1>  
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<p>One of the major results that Waterloo iGEM had accomplished over the summer 2015 is showing that adding restriction sites and a single mutation in the scaffold region of the sgRNA did not affect the functionality of the  sgRNA in CRISPR dCas9 system. The assay used for this experiment is amnis Imaging Flow Cytometry. </p>
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<p>
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In addition, we accomplished to mutate dCas9 at three sites 1135, 1335, and 1337 to alter its PAM site from native NGG to NGAG using the Quick Change protocol. The mutations were confirmed using sequencing. Preliminary experiments were done using sgRNA targeting the J23101 promoter upstream of the GFP using NGAG PAM site. Further result analysis is needed to confirm the functionality of the mutated dCas9. </p>
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{{Waterloo}}
 
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     <h1> Project Results</h1>
 
     <h1> Project Results</h1>
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<p>One of the major results that Waterloo iGEM had accomplished over the summer 2015 is showing that adding restriction sites and a single mutation in the scaffold region of the sgRNA did not affect the functionality of the sgRNA in CRISPR dCas9 system. The assay used for this experiment is amnis Imaging Flow Cytometry. </p>
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    <div class="prevHighlightBox">
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<p>
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        <p>Here you can describe the results of your project and your future plans. </p>
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In addition, we accomplished to mutate dCas9 at three sites 1135, 1335, and 1337 to alter its PAM site from native NGG to NGAG using the Quick Change protocol. The mutations were confirmed using sequencing. Preliminary experiments were done using sgRNA targeting the J23101 promoter upstream of the GFP using NGAG PAM site. Further result analysis is needed to confirm the functionality of the mutated dCas9. </p>
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<p> We were finally able to show that Cas9 was expressed from protoplasts using anti-Cas9 antibodies and dot blots. Additionally, we were able to agroinfiltrate our tentative pCAMBIA + Cas9 + sgRNA vector into <i> Arabidopsis </i>.
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        <h5>What should this page contain?</h5>
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        <ul>
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            <li> Clearly and objectively describe the results of your work.</li>
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            <li> Future plans for the project </li>
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            <li> Considerations for replicating the experiments </li>
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        </ul>
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        <h4> Project Achievements </h4>
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        <p>You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer.</p>
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        <ul>
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            <li>A list of linked bullet points of the successful results during your project</li>
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            <li>A list of linked bullet points of the unsuccessful results during your project. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort.</li>
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        </ul>
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        <h4>Inspiration</h4>
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        <p>See how other teams presented their results.</p>
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        <ul>
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            <li><a href="https://2014.igem.org/Team:TU_Darmstadt/Results/Pathway">2014 TU Darmstadt </a></li>
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            <li><a href="https://2014.igem.org/Team:Imperial/Results">2014 Imperial </a></li>
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            <li><a href="https://2014.igem.org/Team:Paris_Bettencourt/Results">2014 Paris Bettencourt </a></li>
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        </ul>
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    </div>
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</div>
 
</div>
 
</html>
 
</html>
 
{{Waterloo_Footer}}
 
{{Waterloo_Footer}}

Revision as of 03:51, 19 September 2015

Results


Project Results

One of the major results that Waterloo iGEM had accomplished over the summer 2015 is showing that adding restriction sites and a single mutation in the scaffold region of the sgRNA did not affect the functionality of the sgRNA in CRISPR dCas9 system. The assay used for this experiment is amnis Imaging Flow Cytometry.

In addition, we accomplished to mutate dCas9 at three sites 1135, 1335, and 1337 to alter its PAM site from native NGG to NGAG using the Quick Change protocol. The mutations were confirmed using sequencing. Preliminary experiments were done using sgRNA targeting the J23101 promoter upstream of the GFP using NGAG PAM site. Further result analysis is needed to confirm the functionality of the mutated dCas9.

We were finally able to show that Cas9 was expressed from protoplasts using anti-Cas9 antibodies and dot blots. Additionally, we were able to agroinfiltrate our tentative pCAMBIA + Cas9 + sgRNA vector into Arabidopsis .

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