Team:Goettingen/week10-3

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<li>Experiment: <br>We aim to apply directed evolution to reprogram the bacterial chemoreceptor TAR in order to enable the perception of novel substances. A saturated mutagenesis PCR was performed to mutate five individual amino acid residues in the ligand binding site of TAR. The primers that are utilized are the following:<br><br>
<li>Experiment: <br>We aim to apply directed evolution to reprogram the bacterial chemoreceptor TAR in order to enable the perception of novel substances. A saturated mutagenesis PCR was performed to mutate five individual amino acid residues in the ligand binding site of TAR. The primers that are utilized are the following:<br><br>
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TARR6973Fw <br>gcgcaGGAAAGGTCTCACTGAGTnnNTCAGCGGTAnnNATGATGATGGATTCCTCCAATCAACAAAG<br>
+
TARR6973Fw: <br>gcgcaGGAAAGGTCTCACTGAGTnnNTCAGCGGTAnnNATGATGATGGATTCCTCCAATCAACAAAG<br>
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TARR6973Rv <br>gcgcaGAGTAGGTCTCATCAGGTTAATGCGCGTTTGCAG<br>
+
TARR6973Rv: <br>gcgcaGAGTAGGTCTCATCAGGTTAATGCGCGTTTGCAG<br>
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TARY149F150T154Fw <br>gcgcaGGAAAGGTCTCAGGAGCTnnNnnNGCTCAGCCAnnNCAGGGAATGCAAAATGCAATGGGCGAAG<br>
+
TARY149F150T154Fw: <br>gcgcaGGAAAGGTCTCAGGAGCTnnNnnNGCTCAGCCAnnNCAGGGAATGCAAAATGCAATGGGCGAAG<br>
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TARY149F150T154Rv <br>gcgcaGAGTAGGTCTCACTCCAGTATTGCCATAATCTAGGTAATC<br>
+
TARY149F150T154Rv: <br>gcgcaGAGTAGGTCTCACTCCAGTATTGCCATAATCTAGGTAATC<br><br>
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We firstly did a test PCR with both primer pairs to verify the <a href="https://2012.igem.org/Team:Goettingen/Project/Methods">mutagenesis PCR protocol</a> and conditions.
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Revision as of 09:19, 20 September 2012

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