Team:Goettingen/week12-3

From 2012.igem.org

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<b>V07_19_3 2<sup>nd</sup> round: Ligation</b><br>
<b>V07_19_3 2<sup>nd</sup> round: Ligation</b><br>
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<li>Experiment: <br>The PCR was now set up in a volume of 1 mL to generate a high amount of DNA in order to reach the diversity we needed. The reaction was split to 20 50 µL tubes.</li>
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<li>Experiment: <br>The ligation was performed according to the established protocol from <a href="https://2012.igem.org/Team:Goettingen/week10-3">week 10</a>. We set up a volume of 2 mL.</li>
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<h2><b>V07_20 </b></h2><br>
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<b>V07_20_1 2<sup>nd</sup> round: Ethanol precipitation of ligation V07_19 </b><br>
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<ul>
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<li>Experiment: <br>The ethanol precipitaion was performed according to the established protocol from <a href="https://2012.igem.org/Team:Goettingen/week10-3">week 10</a>. </li>
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</ul>
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<br>
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<b>V07_20_2 2<sup>nd</sup> round: Transformation of electrocompetent cells with mutated plasmid mixture</b><br>
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<ul>
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<li>Experiment: <br>The digestion and subsequent clean-up was performed according to the established protocol from <a href="https://2012.igem.org/Team:Goettingen/week10-3">week 10</a>.</li>
</ul>
</ul>
<ul>
<ul>
<li>Observations and results: <br>The corresponding gel showed bands of the expected sizes for each reaction tube. </li>
<li>Observations and results: <br>The corresponding gel showed bands of the expected sizes for each reaction tube. </li>
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</ul>
 +
<br>
 +
<b>V07_20_3 2<sup>nd</sup> round: Ligation</b><br>
 +
<ul>
 +
<li>Experiment: <br>The digestion and subsequent clean-up was performed according to the established protocol from <a href="https://2012.igem.org/Team:Goettingen/week10-3">week 10</a>.</li>
</ul>
</ul>
<br>
<br>

Revision as of 10:21, 24 September 2012

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#3 Chemoreceptor Library - 12th Week

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V07_16


Starpoint of the 2nd round of saturated mutagenesis PCR!
  • Experiment:
    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 PCR protocol is basically the same as for the first round.
  • Observations and results:
    The corresponding agarose gel showed a strong band of the expected size (3769 bp).


V07_17


V07_17_1 Test digestion of Miniprep V07_12
  • Experiment:
    A test digestion using EcoRI and PstI in a 20 µL batch was performed according to protocol to confirm that the mutant plasmid pSB1C3_TAR_QC_18C has actually been isolated correctly.
  • Observations and results:
    The corresponding agarose gel showed two distinctive bands of the expected sizes which led to the conclusion that the plasmid had indeed been isolated!

V07_17_2 2nd round: Saturated mutagenesis PCR, volume 1 mL
  • Experiment:
    The PCR was now set up in a volume of 1 mL to generate a high amount of DNA in order to reach the diversity we needed. The reaction was split to 20 50 µL tubes.


V07_19


V07_19_1 2nd round: PCR clean-up
  • Experiment:
    The clean-up was performed according to the established protocol from week 10.
  • Observations and results:
    The corresponding gel showed bands of the expected sizes for each reaction tube.

V07_19_2 2nd round: Digestion DpnI/BsaI and clean-up
  • Experiment:
    The digestion and subsequent clean-up was performed according to the established protocol from week 10.
  • Observations and results:
    The corresponding gel showed bands of the expected sizes for each reaction tube.

V07_19_3 2nd round: Ligation
  • Experiment:
    The ligation was performed according to the established protocol from week 10. We set up a volume of 2 mL.


V07_20


V07_20_1 2nd round: Ethanol precipitation of ligation V07_19
  • Experiment:
    The ethanol precipitaion was performed according to the established protocol from week 10.

V07_20_2 2nd round: Transformation of electrocompetent cells with mutated plasmid mixture
  • Experiment:
    The digestion and subsequent clean-up was performed according to the established protocol from week 10.
  • Observations and results:
    The corresponding gel showed bands of the expected sizes for each reaction tube.

V07_20_3 2nd round: Ligation
  • Experiment:
    The digestion and subsequent clean-up was performed according to the established protocol from week 10.


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