Team:Goettingen/week15-3

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Revision as of 14:36, 26 September 2012


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V08_06_1 2^nd round: Ethanol precipitation of mutated plasmids

V08_06_2 2^nd round: Transformation of electrocompetent cells with the mutant plasmid mixture

Experiment:
Electrocompetent cells were prepared according to protocol. The transformation was performed according to the established protocol.

V08_07_1 2^nd round: Analysis of transforamtion V08_06

Observations & Results:
Neither the plates nor the liquid culture exhibited bacterial growth. A test gel of the ethano precipitated ligation revealed that, again, our DNA material was lost during the process.

V08_07_2 2^nd round: Mutagenesis PCR was set up again, 1000 µL

V08_08_1 2^nd round: PCR purification

Experiment:
The purification was carried out with the peqGOLD Cycle-pure Kit (PeqLab) following the user manual. Elution in 100 µL EB.

Observations & Results:
Again, we lost a significant amount of DNA material. The subsequent steps (digestion, ligation, transformation) were not carried through because the diversity could not be reached!!

V08_08_2 2^nd round: Mutagenesis PCR was set up again, 1000 µL

V08_09_1 2^nd round: PCR purification of V08_08_2

Experiment:
The purification was performed using the peqGOLD Cycle-pure Kit (PeqLab) following the user manual. Elution in 100 µL EB.

Observations & Results:
Neither the plates nor the liquid culture exhibited bacterial growth. A test gel of the ethano precipitated ligation revealed that, again, our DNA material was lost during the process.

V08_09_2 2^nd round: Mutagenesis PCR was set up again, 1000 µL

V08_10_1 2^nd round: Analysis of transforamtion V08_06

Observations & Results:
Neither the plates nor the liquid culture exhibited bacterial growth. A test gel of the ethano precipitated ligation revealed that, again, our DNA material was lost during the process.

V08_10_2 2^nd round: Mutagenesis PCR was set up again, 1000 µL

@@ Line 62: / Line 62: @@
 <td width="900" bordercolor="black" valign="top">
 <h2><b>V08_08 </b></h2><br>
-<b>V08_08_1 2<sup>nd</sup> round: Analysis of transforamtion V08_06</b><br>
+<b>V08_08_1 2<sup>nd</sup> round: PCR purification</b><br>
 <ul>
-<li>Experiment: <br>Plates and liquid culture were investigated.</li>
+<li>Experiment: <br>The purification was carried out with the peqGOLD Cycle-pure Kit (PeqLab) following the user manual. Elution in 100 µL EB.</li>
 </ul>
 <ul>
-<li>Observations & Results: <br>Neither the plates nor the liquid culture exhibited bacterial growth. A test gel of the ethano precipitated ligation revealed that, again, our DNA material was lost during the process.</li>
+<li>Observations & Results: <br>Again, we lost a significant amount of DNA material. The subsequent steps (digestion, ligation, transformation) were not carried through because the diversity could not be reached!!</li>
 </ul>
 <br>
@@ Line 81: / Line 81: @@
 <td width="900" bordercolor="black" valign="top">
 <h2><b>V08_09 </b></h2><br>
-<b>V08_09_1 2<sup>nd</sup> round: Analysis of transforamtion V08_06</b><br>
+<b>V08_09_1 2<sup>nd</sup> round: PCR purification of V08_08_2</b><br>
 <ul>
-<li>Experiment: <br>Plates and liquid culture were investigated.</li>
+<li>Experiment: <br>The purification was performed using the peqGOLD Cycle-pure Kit (PeqLab) following the user manual. Elution in 100 µL EB.</li>
 </ul>
 <ul>