Team:Yale/Notebook/Week15

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<li><a href="https://2011.igem.org/Team:Yale/Notebook/Week13">Week 13</a></li>
<li><a href="https://2011.igem.org/Team:Yale/Notebook/Week13">Week 13</a></li>
<li><a href="https://2011.igem.org/Team:Yale/Notebook/Week14">Week 14</a></li>
<li><a href="https://2011.igem.org/Team:Yale/Notebook/Week14">Week 14</a></li>
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<li><a id="page" href="https://2011.igem.org/Team:Yale/Notebook/Week15">Week 15</a></li>
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<li><a id="page" href="https://2011.igem.org/Team:Yale/Notebook/Week15-18">Week 15</a></li>
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<h1>Week 15: August 21-28, 2011</h1>
<h1>Week 15: August 21-28, 2011</h1>
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<li> Grew up 6L worth of cells to purify RiAFP, and purified RiAFP. Pellets were flash frozen in liquid nitrogen and stored at -80 C before use. Purification of RiGFP was used for ice recrystallization studies, rat liver morphology studies, and C. elegans survival studies. </li>
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<li> Designed Received primers for PCR amplification of Kanamycin and RiAFP/RiGFP for eventual cross over PCR and integration into the genome.</li>
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<li> Conducted sucessful PCR (insert image). PCR purified sucessful PCR products and treated with DpnI for 30 minutes.</li>
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<li> Crossover PCR was sucessful (insert image) </li>
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<li> Integrated crossover PCR product into genome of EcNR2 strain </li>
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<li> Transformed new EcnR2 strain with plasmid containing T7 polymerase and screened for appropriate colonies. </li>
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<li> Received 22 degenerate oligonucleotide sequences from Keck and performed MAGE. Cultures were grown up to midlog post electroporation and freeze-thawed approximately 14 times before plating. Suriving colonies were isolated, and we will soon put them through another cycle of MAGE. The process will be repeated. </li>
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Revision as of 21:31, 28 September 2011

iGEM Yale

Week 15: August 21-28, 2011

  • Grew up 6L worth of cells to purify RiAFP, and purified RiAFP. Pellets were flash frozen in liquid nitrogen and stored at -80 C before use. Purification of RiGFP was used for ice recrystallization studies, rat liver morphology studies, and C. elegans survival studies.
  • Designed Received primers for PCR amplification of Kanamycin and RiAFP/RiGFP for eventual cross over PCR and integration into the genome.
  • Conducted sucessful PCR (insert image). PCR purified sucessful PCR products and treated with DpnI for 30 minutes.
  • Crossover PCR was sucessful (insert image)
  • Integrated crossover PCR product into genome of EcNR2 strain
  • Transformed new EcnR2 strain with plasmid containing T7 polymerase and screened for appropriate colonies.
  • Received 22 degenerate oligonucleotide sequences from Keck and performed MAGE. Cultures were grown up to midlog post electroporation and freeze-thawed approximately 14 times before plating. Suriving colonies were isolated, and we will soon put them through another cycle of MAGE. The process will be repeated.

    • Retrieved from "http://2011.igem.org/Team:Yale/Notebook/Week15"