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

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	Cut'N'Survive System Functional Principle Sequences Schedule Jump-or-Die System Functional Principle Sequences Schedule ProSearch System Schedule

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==Aim 1: DNA reprod Replication+PCR== PCR1: replication of the Tet-inducible CMV promotor [X]

Primer used: 1TreF 2TreR

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

Primer used: 3TevF 4TevMutEPR

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

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

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) [ ]

Primer used: 15TrecMutSF 16TrecR

PCR8: joining PCR of p14*TEVrecogn [ ]

Primer used: 13TrecF 16TrecR

Aim 2: Assembling Biobricks

Aim 3: Testing products

Aim 4: Testing system

oder

Aim 1: DNA reproduction+PCR

Aim 2: Assembling Biobricks

Aim 3: Testing products

Aim 4: Testing system

alt:

Meilensteine

Für die Projekte haben wir uns jeweils folgende „Meilensteine“ gesetzt:

„TEV-System“

1. Verknüpfen von Bak mit interagierendem Protein A, einem N-Degron und einer Schnittstelle für die TEV-Protesase, sodass Bak noch aktiv bleibt. Davor ist ein induzierbarer Promotor und eine Antibiotikaresistenz geschaltet (Nachweis: induzierbarer Zelltod) Aufwand: ca. 6-8 Wochen, 2000€

2. Verknüpfen von eGFP als Zielgen mit einer TEV-Schnittstelle, einer TEV-Protease, und interagierendem Protein B. (Nachweis: eGFP leuchtet, Proteinchromatographie)

Aufwand: ca. 6-8 Wochen, 2000€, Parallele Arbeit ist möglich und notwendig

3. Einbringen beider Konstrukte in Zellen (Nachweis: eGFP leuchtet, einige Zellen sterben, Proteinchromatographie)

Aufwand: ca. 2 Wochen, 1000€

4. Parallel wird versucht, das erste Konstrukt stabil ins Zellgenom einzubauen.

Aufwand: ca. 2 Wochen, 500€

@@ Line 2: / Line 2: @@
-==Aim 1: DNA reproduction+PCR==
+==Aim 1: DNA reprod
-PCR1: replication of the Tet-inducible CMV promotor
+Replication+PCR==
+PCR1: replication of the Tet-inducible CMV promotor   [X]
 [[Image:PCR1.jpg]]
 Primer used:[[Team:LMU-Munich/Cut'N'survive/Schedule/Primer1| 1TreF]][[Team:LMU-Munich/Cut'N'survive/Schedule/Primer2| 2TreR]]
-PCR2a: replication of the TEVrecogn-NDegron-SF3 Part with mutagenisis (EcoR1 + Pst1)
+PCR2a: replication of the TEVrecogn-NDegron-SF3 Part with mutagenisis (EcoR1 + Pst1) [ ]
+[[Image:PCR2a.jpg]]
-PCR2b: replication of the TEVrecogn-NDegron-SF3 Part with mutagenisis (EcoR1 + Pst1)
+Primer used:[[Team:LMU-Munich/Cut'N'survive/Schedule/Primer3| 3TevF]][[Team:LMU-Munich/Cut'N'survive/Schedule/Primer4| 4TevMutEPR]]
-PCR3: joining PCR of the TEVrecogn-NDegron-SF3 Part
+PCR2b: replication of the TEVrecogn-NDegron-SF3 Part with mutagenisis (EcoR1 + Pst1) [ ]
+[[Image:PCR2b.jpg]]
-PCR4a: replication human bak with mutagenisis (Pst1)
+Primer used:[[Team:LMU-Munich/Cut'N'survive/Schedule/Primer5| 5TevMutEPF]][[Team:LMU-Munich/Cut'N'survive/Schedule/Primer6| 6TevR]]
-PCR4b: replication human bak with mutagenisis (Pst1)
+PCR3: joining PCR of the TEVrecogn-NDegron-SF3 Part [ ]
+[[Image:PCR3.jpg]]
-PCR5: joining PCR of human bak
+Primer used:[[Team:LMU-Munich/Cut'N'survive/Schedule/Primer3| 3TevF]][[Team:LMU-Munich/Cut'N'survive/Schedule/Primer6| 6TevR]]
-PCR6: replication of the SV40-polyadenylation site
+PCR4a: replication human bak with mutagenisis (Pst1) [ ]
+[[Image:PCR4a.jpg]]
-PCR7a: replication of the p14*TEVrecogn with mutagenisis (Spe1)
+Primer used:[[Team:LMU-Munich/Cut'N'survive/Schedule/Primer7| 7BakF]][[Team:LMU-Munich/Cut'N'survive/Schedule/Primer8| 8BakMutPR]]
-PCR7b: replication of the p14*TEVrecogn with mutagenisis (Spe1)
+PCR4b: replication human bak with mutagenisis (Pst1) [ ]
+[[Image:PCR4b.jpg]]
-PCR8: joining PCR of p14*TEVrecogn
+Primer used:[[Team:LMU-Munich/Cut'N'survive/Schedule/Primer9| 9BakMutPF]][[Team:LMU-Munich/Cut'N'survive/Schedule/Primer10| 10BakR]]
+PCR5: joining PCR of human bak [ ]
+[[Image:PCR5.jpg]]
+Primer used:[[Team:LMU-Munich/Cut'N'survive/Schedule/Primer7| 7BakF]][[Team:LMU-Munich/Cut'N'survive/Schedule/Primer10| 10BakR]]
+PCR6: replication of the SV40-polyadenylation site [X]
+[[Image:PCR6.jpg]]
+Primer used:[[Team:LMU-Munich/Cut'N'survive/Schedule/Primer11| 11PAF]][[Team:LMU-Munich/Cut'N'survive/Schedule/Primer12| 12PAR]]
+PCR7a: replication of the p14*TEVrecogn with mutagenisis (Spe1) [ ]
+[[Image:PCR7a.jpg]]
+Primer used:[[Team:LMU-Munich/Cut'N'survive/Schedule/Primer13| 13TrecF]][[Team:LMU-Munich/Cut'N'survive/Schedule/Primer14| 14TrecMutSR]]
+PCR7b: replication of the p14*TEVrecogn with mutagenisis (Spe1) [ ]
+[[Image:PCR7b.jpg]]
+Primer used:[[Team:LMU-Munich/Cut'N'survive/Schedule/Primer15| 15TrecMutSF]][[Team:LMU-Munich/Cut'N'survive/Schedule/Primer16| 16TrecR]]
+PCR8: joining PCR of p14*TEVrecogn [ ]
+[[Image:PCR8.jpg]]
+Primer used:[[Team:LMU-Munich/Cut'N'survive/Schedule/Primer13| 13TrecF]][[Team:LMU-Munich/Cut'N'survive/Schedule/Primer16| 16TrecR]]
 ==Aim 2: Assembling Biobricks==