Difference between revisions of "Team:Paris Bettencourt/Notebook/Phytase"
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<h3>Gene PHO85</h3> | <h3>Gene PHO85</h3> | ||
− | 5’Primer of | + | 5’Primer of Kanamycin resistance gene with tails using to transformation with the PHO85 gene of the yeast. |
<br>5’-<span style="color:#9A1717">TATCATTATATATACATGGCTACGGTTTTTCGCTGACGGGCTGCG</span><span style="color:#179A89">ATAATCATTTGCA | <br>5’-<span style="color:#9A1717">TATCATTATATATACATGGCTACGGTTTTTCGCTGACGGGCTGCG</span><span style="color:#179A89">ATAATCATTTGCA | ||
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<br> | <br> | ||
− | <br>3’Primer of | + | <br>3’Primer of Kanamycin resistance gene with tails using to transformation with the PHO85 gene of the yeast. |
<br>3’-<span style="color:#9A1717">AAGGGATATATAGCGCGGCAAACTGGGCAAACTTGAGCAATACCA</span><span style="color:#179A89">CAGCAGTATAG | <br>3’-<span style="color:#9A1717">AAGGGATATATAGCGCGGCAAACTGGGCAAACTTGAGCAATACCA</span><span style="color:#179A89">CAGCAGTATAG | ||
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<br> | <br> | ||
− | <br><span style="color:#9A1717"> - tail | + | <br><span style="color:#9A1717"> - tail homology PHO85 </span> |
− | <br><span style="color:#179A89"> - Primer | + | <br><span style="color:#179A89"> - Primer Kanamycin </span><br> |
<br><h3>Gene PHO80</h3> | <br><h3>Gene PHO80</h3> | ||
− | 5’Primer of | + | 5’Primer of <i>Kanamycin<i> resistance gene with tails using to transformation with the PHO80 gene of the yeast. |
<br>5’-<span style="color:#9A1717">ATCATAAGACGAGGATATCCTTTGGAGACTCATAGAAATC</span><span style="color:#179A89">ATAATCATTTGCATCCAT | <br>5’-<span style="color:#9A1717">ATCATAAGACGAGGATATCCTTTGGAGACTCATAGAAATC</span><span style="color:#179A89">ATAATCATTTGCATCCAT | ||
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− | <br>3’Primer of | + | <br>3’Primer of Kanamycin resistance gene with tails using to transformation with the PHO80 gene of the yeast. |
<br>3’-<span style="color:#9A1717">CTCAATCATGATTGCTTTCATAATACCCCACGAAAAATCA</span><span style="color:#179A89">CAGCAGTATAGCGACCA | <br>3’-<span style="color:#9A1717">CTCAATCATGATTGCTTTCATAATACCCCACGAAAAATCA</span><span style="color:#179A89">CAGCAGTATAGCGACCA | ||
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<br> | <br> | ||
− | <br><span style="color:#9A1717"> - tail | + | <br><span style="color:#9A1717"> - tail homology PHO80 </span> |
− | <br><span style="color:#179A89"> - Primer | + | <br><span style="color:#179A89"> - Primer Kanamycin </span><br> |
<br><h3>Gene FRT + PHO85</h3> | <br><h3>Gene FRT + PHO85</h3> | ||
− | 5’Primer of | + | 5’Primer of Kanamycin resistance gene with tails using to transformation with the PHO85 gene of the yeast, including FRT sequence to delete both of PHO80 and PHO85. |
<br>5’-<span style="color:#9A1717">TATCATTATATATACATGGCTACGGTTTTTCGCTGACGGGCTGCG</span><span style="color:#36C40F">GAAGTTCCTATTC | <br>5’-<span style="color:#9A1717">TATCATTATATATACATGGCTACGGTTTTTCGCTGACGGGCTGCG</span><span style="color:#36C40F">GAAGTTCCTATTC | ||
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<br> | <br> | ||
− | <br>3’Primer of | + | <br>3’Primer of Kanamycin resistance gene with tails using to transformation with the PHO85 gene of the yeast, including FRT sequence to delete both of PHO80 and PHO85. |
<br>3’-<span style="color:#9A1717">AAGGGATATATAGCGCGGCAAACTGGGCAAACTTGAGCAATACCA</span><span style="color:#36C40F">CTTCAAGGATAT | <br>3’-<span style="color:#9A1717">AAGGGATATATAGCGCGGCAAACTGGGCAAACTTGAGCAATACCA</span><span style="color:#36C40F">CTTCAAGGATAT | ||
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<br>GAAAGATCTCTTATCCTTGAAG</span><span style="color:#179A89">CAGCAGTATAGCGACCAGCATTC</span>-5’ | <br>GAAAGATCTCTTATCCTTGAAG</span><span style="color:#179A89">CAGCAGTATAGCGACCAGCATTC</span>-5’ | ||
<br> | <br> | ||
− | <br><span style="color:#9A1717"> - tail | + | <br><span style="color:#9A1717"> - tail homology PHO85 </span> |
<br><span style="color:#36C40F"> - FRT</span> | <br><span style="color:#36C40F"> - FRT</span> | ||
− | <br><span style="color:#179A89"> - Primer | + | <br><span style="color:#179A89"> - Primer Kanamycin </span><br> |
<br><h1>12/08/15</h1> | <br><h1>12/08/15</h1> | ||
<h2>Culture</h2> | <h2>Culture</h2> | ||
− | The | + | The Saccharomyces cerevisiae SK1 was thaw and sow 100µL of it on YPD medium overnight. (at 30°C) |
<br>This yeast will be transformed.<br> | <br>This yeast will be transformed.<br> | ||
<br><h2>PCR</h2> | <br><h2>PCR</h2> |
Revision as of 20:27, 14 August 2015