Team:LMU-Munich/Jump-or-die/Schedule

From 2010.igem.org

(Difference between revisions)
(Construct 1)
(Construct 2)
Line 91: Line 91:
=== Construct 2 ===
=== Construct 2 ===
-
- transform into HeLa cells and see if eGFP is being translated
+
[[Image: Jump12.jpg | 400pxs | Jump construct 2]]
-
-> Check if CMV-promoter is working and construct is being read-off completely
+
- sequencing
-
 
+
-
- Western Blot
+
-
FALSEEE
+
==Aim 4: Testing system==
==Aim 4: Testing system==
{{:Team:LMU-Munich/Templates/Page Footer}}
{{:Team:LMU-Munich/Templates/Page Footer}}

Revision as of 10:49, 30 August 2010


Contents

Aim 1: DNA reproduction+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 product size: 492 bp


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

Primer used: 7BakF 8BakMutPR expected product size: 330 bp


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

Primer used: 9BakMutPF 10BakR expected product size: 376 bp


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

Primer used: 7BakF 10BakR expected product size: 688 bp


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

Primer used: 11PAF 12PAR expected product size: 237 bp


PCR9: replication of eGFP with attP in primer [ ] PCR9.jpg

Primer used: 20eGFPattPF 21eGFPR expected product size: 808 bp


PCR10: Replikation of PhiC31o [ ] PCR10.jpg

Primer used: 22PC31oF 23PC31oR expected product size: 1888 bp

Note: for attB two Primers ( 18attBF 19attBR) used.

Aim 2: Assembling Biobricks

We are using the 3A System to assemble Biobricks.

Assembling Construct 1

J1.jpg

Assembling Construct 2

J2.jpg

Assembling Construct 3

J3.jpg

Aim 3: Testing products

Construct 1

Jump construct 1

- transform into HeLa cells to see if they survive.

-> Check the leakiness of tet-on-promoter

- induce tet-on-promoter to see, if cells die.

-> Check if construct 1 is working

Construct 2

Jump construct 2

- sequencing

Aim 4: Testing system