Difference between revisions of "Team:ASIJ Tokyo/Experiments"

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<h1 id="header"> EXPERIMENTS </h1>
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                <h1 id="header"> EXPERIMENTS </h1>
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    <h4><a href="https://static.igem.org/mediawiki/2018/c/cc/T--ASIJ_Tokyo--crisprpart1.pdf" target="none">Part I  CRISPR</a></h3>
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<br> <h4><a href="https://static.igem.org/mediawiki/2018/3/30/T--ASIJ_Tokyo--crisprpart2.pdf">Part II CRISPR Confirmation (genotypic)</a></h3>
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<br><h4>Part III: Designing of the new constructs (this is just modeling the construct design and not really a protocol)</h3>
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<br> <h4><a href="https://static.igem.org/mediawiki/2018/f/fd/T--ASIJ_Tokyo--gibson.pdf">Part IV:  Gibson Assembly of Constructs</a></h3>
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<br> <h4>Part V: Deciding which fluorescent marker to use (water melon tubes, and plates)..deciding then to only use the GFP ones</h3>
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<br> <h4>Part VI: Genotypic and Phenotypic Proof of Concept</h3>
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    <ul>
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      <li>a. Agarose gel electrophoresis of plasmid constructs (agarose gel electorphoresis protocol)</li>
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        <li>b. Phenotypic Proof (Transformation and growing on selection plates)</li>
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    </ul>
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    <h4>Part VII:  Quantifying Protein</h3>
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    <ul>
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        <li>a.  <a href="https://static.igem.org/mediawiki/2018/a/a7/T--ASIJ_Tokyo--oggfp.pdf">GFP protein purification using  column chromatography</a></li>
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        <li>b.  <a href="https://static.igem.org/mediawiki/2018/5/56/T--ASIJ_Tokyo--ogsialic.pdf">Sialic Acid Assay - original version of protocol </a></li>
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    </ul>
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<h4>Part VIII: Incorporation of New information to use His-tag</h3>
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<ul>
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    <li>a. Redesign construct to include His-tag</li>
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    <li>b. Gibson assembly </li>
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    <li>c. Transformation  </li>
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    <li>d. <a href="https://static.igem.org/mediawiki/2018/2/2b/T--ASIJ_Tokyo--histag.pdf">Protein Purification using His Tag Protein Chromatography -
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        redone  Oct 10  (10 trials)</a></li>
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    <li>c. <a href="https://static.igem.org/mediawiki/2018/8/84/T--ASIJ_Tokyo--esla.pdf">Sialic Acid Purification redone using new protocol (6 trials) </a></li>
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    <li>d. Fluoroscence Microplate Reading to Quantify Protein Concentration of Constructs
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        using GFP standard curve </li>
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    <li>e. Fluorescence Microplate Reading to Quantify Sialic Acid Concentration
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        using Sialic Acid Standard Curve </li>
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    <li>f. <a href="https://static.igem.org/mediawiki/2018/4/44/T--ASIJ_Tokyo--electro.pdf">Final Polyacrylamide Gel Confirmation of Constructs </a></li>
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Revision as of 15:02, 14 October 2018

EXPERIMENTS

Part I CRISPR


Part II CRISPR Confirmation (genotypic)


Part III: Designing of the new constructs (this is just modeling the construct design and not really a protocol)


Part IV: Gibson Assembly of Constructs


Part V: Deciding which fluorescent marker to use (water melon tubes, and plates)..deciding then to only use the GFP ones


Part VI: Genotypic and Phenotypic Proof of Concept

  • a. Agarose gel electrophoresis of plasmid constructs (agarose gel electorphoresis protocol)
  • b. Phenotypic Proof (Transformation and growing on selection plates)

Part VII: Quantifying Protein

Part VIII: Incorporation of New information to use His-tag