Team:Goettingen/week17-3

From 2012.igem.org

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<h2><b>V08_20 </b></h2><br>
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<h2><b>V08_21 </b></h2><br>
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<b>V08_20_1 Attempt b: increasing the library diversity!</b><br>
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<b>V08_21_1 Attempt b: <i>Dpn</i>I/<i>Bsa</i>I digest and subsequent purification</b><br>
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<li>Experiment: <br>A PCR (1000 µL) using the 2<sup>nd</sup> round primers (Tar6973 for and Tar6973 rev) and the liquid culture pellet Miniprep that was used at <a href="https://2012.igem.org/Team:Goettingen/week13-3">V07_25</a> as template was performed. Protocol according to <a href="https://2012.igem.org/Team:Goettingen/week10-3">week 10</a>. By doing this and later combine the products, the diversity can be increased. Purification was carried our using the peqGOLD Cycle-pure Kit (PeqLab) with Miniprep Kit columns following the user manual.</li>
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<li>Experiment: <br>The digest and purification was performed following the protocol from <a href="https://2012.igem.org/Team:Goettingen/week10-3">week 10</a>.</li>
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<b>V08_20_2 2<sup>nd</sup> round: Plasmid preparation from pellets #1-#4 V08_17 </b><br>
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<b>V08_21_2 Attempt b: Ligation, 2000 µL </b><br>
<ul>
<ul>
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<li>Experiment: <br>peqGOLD Xchange Plasmid Midi Kit (PeqLab) was used following the user manual. Elution in 100 µL H<sub>2</sub>O. Stored at -20 °C.</li>
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<li>Experiment: <br>The ligation was performed according to the protocol from <a href="https://2012.igem.org/Team:Goettingen/week10-3">week 10</a>.</li>
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<h2><b>V08_22 </b></h2><br>
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<b>V08_22_1 Attempt b: <i></b><br>
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<ul>
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<li>Experiment: <br>The digest and purification was performed following the 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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<ul>
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<li>Observations & Results: <br>The corresponding agarose gel showed a band of the expected size.</li>
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<br>
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<b>V08_22_2 Attempt b: Ligation, 2000 µL </b><br>
<ul>
<ul>
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<li>Observations & Results: <br>DNA concentrations were determined as follows using nanodrop:<br>
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<li>Experiment: <br>The ligation was performed according to the protocol from <a href="https://2012.igem.org/Team:Goettingen/week10-3">week 10</a>.</li>
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#1 - 521 ng/µL<br>
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#2 - 1035 ng/µL<br>
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#3 - 2145 ng/µL<br>
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#4 - 1103 ng/µL<br>
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Revision as of 20:12, 26 September 2012

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#3 Chemoreceptor Library - 17th Week

Back to overview

V08_20


V08_20_1 Attempt b: increasing the library diversity!
  • Experiment:
    A PCR (1000 µL) using the 2nd round primers (Tar6973 for and Tar6973 rev) and the liquid culture pellet Miniprep that was used at V07_25 as template was performed. Protocol according to week 10. By doing this and later combine the products, the diversity can be increased. Purification was carried our using the peqGOLD Cycle-pure Kit (PeqLab) with Miniprep Kit columns following the user manual.
  • Observations & Results:
    The corresponding agarose gel showed a band of the expected size.

V08_20_2 2nd round: Plasmid preparation from pellets #1-#4 V08_17
  • Experiment:
    peqGOLD Xchange Plasmid Midi Kit (PeqLab) was used following the user manual. Elution in 100 µL H2O. Stored at -20 °C.
  • Observations & Results:
    DNA concentrations were determined as follows using nanodrop:
    #1 - 521 ng/µL
    #2 - 1035 ng/µL
    #3 - 2145 ng/µL
    #4 - 1103 ng/µL


V08_21


V08_21_1 Attempt b: DpnI/BsaI digest and subsequent purification
  • Experiment:
    The digest and purification was performed following the protocol from week 10.
  • Observations & Results:
    The corresponding agarose gel showed a band of the expected size.

V08_21_2 Attempt b: Ligation, 2000 µL
  • Experiment:
    The ligation was performed according to the protocol from week 10.


V08_22


V08_22_1 Attempt b:
  • Experiment:
    The digest and purification was performed following the protocol from week 10.
  • Observations & Results:
    The corresponding agarose gel showed a band of the expected size.

V08_22_2 Attempt b: Ligation, 2000 µL
  • Experiment:
    The ligation was performed according to the protocol from week 10.


Back to overview
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