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Aim 1: DNA replication+PCR
For PCR key see PCR key
Colourcode for the primers: Annealing part, mutagenised part, other sequences integrated into primer
PCR1: replication of the Tet-inducible CMV promotor [X]
Primer used: 1TreF 2TreR expected productsize: 492bp
PCR2a: replication of the TEVrecogn-NDegron-SF3 Part with mutagenisis (EcoR1 + Pst1) [X]
Primer used: 3TevF 4TevMutEPR
PCR2b: replication of the TEVrecogn-NDegron-SF3 Part with mutagenisis (EcoR1 + Pst1) [X]
Primer used: 5TevMutEPF 6TevR
PCR3: joining PCR of the TEVrecogn-NDegron-SF3 Part [ ]
Primer used: 3TevF 6TevR
PCR4a: replication human bak with mutagenisis (Pst1) [ ]
Primer used: 7BakF 8BakMutPR
PCR4b: replication human bak with mutagenisis (Pst1) [ ]
Primer used: 9BakMutPF 10BakR
PCR5: joining PCR of human bak [ ]
Primer used: 7BakF 10BakR
expected product size: 685 bp
PCR6: replication of the SV40-polyadenylation site [X]
Primer used: 11PAF 12PAR
PCR7a: replication of the p14*TEVrecogn with mutagenisis (Spe1) [ ]
Primer used: 13TrecF 14TrecMutSR
PCR7b: replication of the p14*TEVrecogn with mutagenisis (Spe1) with TEV recogn in primer (16TrecR) [ ]
Primer used: 15TrecMutSF 16TrecR
PCR8: joining PCR of p14*TEVrecogn with TEV recogn in primer (16TrecR) [ ]
Primer used: 13TrecF 16TrecR
Aim 2: Assembling Biobricks
Aim 3: Testing products
Aim 4: Testing system
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