Team:LMU-Munich/Cut'N'survive/Schedule

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Contents

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] PCR1.jpg

Primer used: 1TreF 2TreR expected productsize: 492bp

PCR2a: replication of the TEVrecogn-NDegron-SF3 Part with mutagenisis (EcoR1 + Pst1) [X] PCR2a.jpg

Primer used: 3TevF 4TevMutEPR

PCR2b: replication of the TEVrecogn-NDegron-SF3 Part with mutagenisis (EcoR1 + Pst1) [X] PCR2b.jpg

Primer used: 5TevMutEPF 6TevR

PCR3: joining PCR of the TEVrecogn-NDegron-SF3 Part [ ] PCR3.jpg

Primer used: 3TevF 6TevR

PCR4a: replication human bak with mutagenisis (Pst1) [ ] PCR4a.jpg

Primer used: 7BakF 8BakMutPR

PCR4b: replication human bak with mutagenisis (Pst1) [ ] PCR4b.jpg

Primer used: 9BakMutPF 10BakR

PCR5: joining PCR of human bak [ ] PCR5.jpg

Primer used: 7BakF 10BakR

expected product size: 685 bp

PCR6: replication of the SV40-polyadenylation site [X] PCR6.jpg

Primer used: 11PAF 12PAR

PCR7a: replication of the p14*TEVrecogn with mutagenisis (Spe1) [ ] PCR7a.jpg

Primer used: 13TrecF 14TrecMutSR

PCR7b: replication of the p14*TEVrecogn with mutagenisis (Spe1) with TEV recogn in primer (16TrecR) [ ] PCR7b.jpg

Primer used: 15TrecMutSF 16TrecR

PCR8: joining PCR of p14*TEVrecogn with TEV recogn in primer (16TrecR) [ ] PCR8.jpg

Primer used: 13TrecF 16TrecR

Aim 2: Assembling Biobricks

Aim 3: Testing products

Aim 4: Testing system