Team:Goettingen/week12-3

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Revision as of 18:57, 22 September 2012


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Starpoint of the 2^nd 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_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 2^nd 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.

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-<b>V07_17_2 2<sup>nd</sup> round: saturated mutagenesis PCR, volume 1 mL</b><br>
+<b>V07_17_2 2<sup>nd</sup> round: Saturated mutagenesis PCR, volume 1 mL</b><br>
 <ul>
 <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. </li>