Team:Goettingen/week12-3
From 2012.igem.org
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<b>Starpoint of the 2<sup>nd</sup> round of saturated mutagenesis PCR!</b><br> | <b>Starpoint of the 2<sup>nd</sup> round of saturated mutagenesis PCR!</b><br> | ||
<ul> | <ul> | ||
- | <li>Experiment: <br>For the second round of saturated mutagenesis PCR we used the primers Tar6973Fw and Tar6973Rv. Here, aa residues 69 and 73 will be mutated. The <a href="https://2012.igem.org/Team:Goettingen/Project/Methods">PCR protocol</a> is basically the same as for the first round. </li> | + | <li>Experiment: <br>For the second round of saturated mutagenesis PCR we used the primers Tar6973Fw and Tar6973Rv. Here, aa residues 69 and 73 will be mutated. We used the Miniprep <a href="https://2012.igem.org/Team:Goettingen/week11-3">V07_12</a> of the liquid culture from the first mutagenesis round. It contains multiple plasmids that are mutated for the first three aa sites. The <a href="https://2012.igem.org/Team:Goettingen/Project/Methods">PCR protocol</a> is basically the same as for the first round. </li> |
</ul> | </ul> | ||
<ul> | <ul> | ||
- | <li>Observations | + | <li>Observations & Results: <br>The corresponding agarose gel showed a strong band of the expected size (3769 bp). </li> |
</ul> | </ul> | ||
<br></td></tr> | <br></td></tr> | ||
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</ul> | </ul> | ||
<ul> | <ul> | ||
- | <li>Observations | + | <li>Observations & Results: <br>The corresponding agarose gel showed two distinctive bands of the expected sizes which led to the conclusion that the plasmid had indeed been isolated! </li> |
</ul> | </ul> | ||
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</ul> | </ul> | ||
<ul> | <ul> | ||
- | <li>Observations | + | <li>Observations & Results: <br>The corresponding gel showed bands of the expected sizes for each reaction tube. </li> |
</ul> | </ul> | ||
<br> | <br> | ||
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</ul> | </ul> | ||
<ul> | <ul> | ||
- | <li>Observations | + | <li>Observations & Results: <br>The corresponding gel showed bands of the expected sizes for each reaction tube. </li> |
</ul> | </ul> | ||
<br> | <br> | ||
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<b>V07_20_2 2<sup>nd</sup> round: Transformation of electrocompetent cells with mutated plasmid mixture</b><br> | <b>V07_20_2 2<sup>nd</sup> round: Transformation of electrocompetent cells with mutated plasmid mixture</b><br> | ||
<ul> | <ul> | ||
- | <li>Experiment: <br>The preparation of electrocompetent cells and the subsequent transformation were performed according to the established protocol from <a href="https://2012.igem.org/Team:Goettingen/week10-3">week 10</a>. | + | <li>Experiment: <br>The preparation of electrocompetent cells and the subsequent transformation were performed according to the established protocol from <a href="https://2012.igem.org/Team:Goettingen/week10-3">week 10</a>. We adjusted the volume of liquid culture to 250 mL to reach diversity.</li> |
- | + | </ul><br> | |
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- | </ul> | + | |
- | <br> | + | |
</td></tr> | </td></tr> | ||
</table> | </table> |
Latest revision as of 13:33, 25 September 2012
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#3 Chemoreceptor Library - 12th WeekBack to overview
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