PCR and mutagenisis of the digestion sites
Primer table : Primer sequences, intern numeration, Tm
PCR1a: replication of Pdu D with mutagenesis [ ]
Primer used: 1P1D, mpduD rev expected product size: bp
PCR1b: replication of Pdu D with mutagenesis [ ]
Primer used: mpduD fwd, 3P2D expected product size: bp
PCR2: Joining PCR of PduD [ ]
Primer used: 1P1D, 3P2D expected product size: bp
PCR3a: replication of Pdu C with mutagenesis [ ]
Primer used: 2P1C, mpduC rev expected product size: bp
PCR3b: replication of Pdu C with mutagenesis [ ]
Primer used: mpduC fwd, 4P2C expected product size: bp
PCR4: Joining PCR of Pdu C [ ]
Primer used: 2P1C, 4P2C expected product size: bp
PCR5: replication of AurF to be joined with Pdu D [ ]
Primer used: 5P3AD, 9P4A expected product size: bp
PCR6a: replication of LctO with mutagenesis [ ]
Primer used: 8P3LC, m1lctO rev expected product size: bp
PCR6b: replication of LctO with mutagenesis [ ]
Primer used: m1lctO fwd, 10P4L expected product size: bp
PCR7: Joining PCR of LctO [ ]
Primer used: 8P3LC, 10P4L expected product size: bp
PCR8a: replication of LctO with mutagenesis [ ]
Primer used: 8P3LC, m2lctO rev expected product size: bp
PCR8b: replication of LctO with mutagenesis [ ]
Primer used: m2lctO fwd, 10P4L expected product size: bp
PCR9: Joining PCR of LctO [ ]
Primer used: 8P3LC, 10P4L expected product size: bp
PCR10: Joining PCR of AurF and PduD [ ]
Primer used: 1P1D, 9P4A expected product size: bp
PCR11: Joining PCR of LctO and PduC [ ]
Primer used: 2P1C, 10P4L expected product size: bp
PCR12a: replication of the Pdu-Operon with mutagenesis [ ]
Primer used: pduA fwd, mpduA rev expected product size: bp
PCR12b: replication of the Pdu-Operon with mutagenesis [ ]
Primer used: mpduA fwd,pduU rev expected product size: bp
PCR13: Joining PCR of the Pdu-Operon [ ]
Primer used: pduA fwd,pduU rev expected product size: bp
PCR14a: replication of the Pdu-Operon with mutagenesis [ ]
Primer used: pduA fwd,mpduB rev expected product size: bp
PCR14b: replication of the Pdu-Operon with mutagenesis [ ]
Primer used: mpduB fwd,pduU rev expected product size: bp
PCR15: Joining PCR of the Pdu-Operon [ ]
Primer used:pduA fwd,pduU rev expected product size: bp
PCR16a: replication of the Pdu-Operon with mutagenesis [ ]
Primer used:pduA fwd,mpduJ rev expected product size: bp
PCR16b: replication of the Pdu-Operon with mutagenesis [ ]
Primer used:mpduJ fwd,pduU rev expected product size: bp
PCR17: Joining PCR of the Pdu-Operon [ ]
Primer used:pduA fwd,pduU rev expected product size: bp
In Vitro testing of the chemical kinetics with the enzymes
Transforming the construct in bacteria
Testing the system
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