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Team:LMU-Munich/Cut'N'survive/Schedule
From 2010.igem.org
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==Aim 1: DNA reproduction+PCR== | ==Aim 1: DNA reproduction+PCR== | ||
PCR1: replication of the Tet-inducible CMV promotor | PCR1: replication of the Tet-inducible CMV promotor | ||
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PCR2a: replication of the TEVrecogn-NDegron-SF3 Part with mutagenisis (EcoR1 + Pst1) | PCR2a: replication of the TEVrecogn-NDegron-SF3 Part with mutagenisis (EcoR1 + Pst1) | ||
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PCR2b: replication of the TEVrecogn-NDegron-SF3 Part with mutagenisis (EcoR1 + Pst1) | PCR2b: replication of the TEVrecogn-NDegron-SF3 Part with mutagenisis (EcoR1 + Pst1) | ||
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PCR3: joining PCR of the TEVrecogn-NDegron-SF3 Part | PCR3: joining PCR of the TEVrecogn-NDegron-SF3 Part | ||
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PCR4a: replication human bak with mutagenisis (Pst1) | PCR4a: replication human bak with mutagenisis (Pst1) | ||
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PCR4b: replication human bak with mutagenisis (Pst1) | PCR4b: replication human bak with mutagenisis (Pst1) | ||
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PCR5: joining PCR of human bak | PCR5: joining PCR of human bak | ||
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PCR6: replication of the SV40-polyadenylation site | PCR6: replication of the SV40-polyadenylation site | ||
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PCR7a: replication of the p14*TEVrecogn with mutagenisis (Spe1) | PCR7a: replication of the p14*TEVrecogn with mutagenisis (Spe1) | ||
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PCR7b: replication of the p14*TEVrecogn with mutagenisis (Spe1) | PCR7b: replication of the p14*TEVrecogn with mutagenisis (Spe1) | ||
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PCR8: joining PCR of p14*TEVrecogn | PCR8: joining PCR of p14*TEVrecogn | ||
Revision as of 10:30, 18 August 2010
PCR1: replication of the Tet-inducible CMV promotor
PCR2a: replication of the TEVrecogn-NDegron-SF3 Part with mutagenisis (EcoR1 + Pst1)
PCR2b: replication of the TEVrecogn-NDegron-SF3 Part with mutagenisis (EcoR1 + Pst1)
PCR3: joining PCR of the TEVrecogn-NDegron-SF3 Part
PCR4a: replication human bak with mutagenisis (Pst1)
PCR4b: replication human bak with mutagenisis (Pst1)
PCR5: joining PCR of human bak
PCR6: replication of the SV40-polyadenylation site
PCR7a: replication of the p14*TEVrecogn with mutagenisis (Spe1)
PCR7b: replication of the p14*TEVrecogn with mutagenisis (Spe1)
PCR8: joining PCR of p14*TEVrecogn
oder
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€
Contents
Aim 1: DNA reproduction+PCR
Aim 2: Assembling Biobricks
Aim 3: Testing products
Aim 4: Testing system
