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

From 2010.igem.org

(Difference between revisions)
(Aim 1: DNA reproduction+PCR)
(Aim 1: DNA reproduction+PCR)
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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
 +
PCR2a: replication of the TEVrecogn-NDegron-SF3 Part with mutagenisis (EcoR1 + Pst1)
PCR2a: replication of the TEVrecogn-NDegron-SF3 Part with mutagenisis (EcoR1 + Pst1)
 +
PCR2b: 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
PCR3: joining PCR of the TEVrecogn-NDegron-SF3 Part
 +
PCR4a: replication human bak with mutagenisis (Pst1)
PCR4a: replication human bak with mutagenisis (Pst1)
 +
PCR4b: replication human bak with mutagenisis (Pst1)
PCR4b: replication human bak with mutagenisis (Pst1)
 +
PCR5: joining PCR of human bak
PCR5: joining PCR of human bak
 +
PCR6: replication of the SV40-polyadenylation site
PCR6: replication of the SV40-polyadenylation site
 +
PCR7a: replication of the p14*TEVrecogn with mutagenisis (Spe1)
PCR7a: replication of the p14*TEVrecogn with mutagenisis (Spe1)
 +
PCR7b: replication of the p14*TEVrecogn with mutagenisis (Spe1)
PCR7b: replication of the p14*TEVrecogn with mutagenisis (Spe1)
 +
PCR8: joining PCR of p14*TEVrecogn
PCR8: joining PCR of p14*TEVrecogn

Revision as of 10:30, 18 August 2010



Contents

Aim 1: DNA reproduction+PCR

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

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€