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Team:LMU-Munich/Jump-or-die/Schedule
From 2010.igem.org
(→Aim 2: Inserting PCR Products in pSB1C3 and verifying Sequence) |
(→Aim 4: Testing products) |
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| - | [[Image: | + | [[Image: Konstruktjump1.PNG | 400pxs | Jump construct 1]] |
- Transform into HeLa cells to see if they survive. | - Transform into HeLa cells to see if they survive. | ||
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=== Construct 2 === | === Construct 2 === | ||
| - | [[Image: | + | [[Image: Konstruktjump2.PNG | 400pxs | Jump construct 2]] |
- Sequencing | - Sequencing | ||
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=== Construct 3 === | === Construct 3 === | ||
| - | [[Image: | + | [[Image: Konstruktjump3.PNG | 400pxs | Jump construct 3]] |
- Transform into HeLa cells | - Transform into HeLa cells | ||
Revision as of 19:12, 24 October 2010
For PCR key see PCR key
Colourcode for the primers: Annealing part, mutagenised part, other sequences integrated into primer
Primer used: 1TreF 2TreR
expected product size: 492 bp
Primer used: 7BakF 8BakMutPR
expected product size: 330 bp
Primer used: 9BakMutPF 10BakR
expected product size: 376 bp
Primer used: 7BakF 10BakR
expected product size: 688 bp
Primer used: 11PAF 12PAR
expected product size: 237 bp
Primer used: 20eGFPattPF 21eGFPR
expected product size: 808 bp
Primer used: 22PC31oF 23PC31oR
expected product size: 1888 bp
Note: for attB two Primers ( 18attBF 19attBR) used.
PCR1 (BBa_K368001): Insertion [ ] -> Sequence of PCR1 from sequencing: confirmed [ ]
PCR5 (BBa_K368017): Insertion [ ] -> Sequence of PCR5 from sequencing: confirmed [ ]
PCR6 (BBa_K368018): Insertion [ ] -> Sequence of PCR6 from sequencing: confirmed [ ]
PCR9 (BBa_K368021): Insertion [ ] -> Sequence of PCR9 from sequencing: confirmed [ ]
PCR10 (BBa_K368022): Insertion [ ] -> Sequence of PCR10 from sequencing: confirmed [ ]
We are using the 3A System to assemble Biobricks.
Assembly of BioBricks:
-> Sequence of BB1 from sequencing: confirmed [ ]
-> Sequence of BB2 from sequencing: confirmed [ ]
-> Sequence of BB3 from sequencing: confirmed [ ]
-> Sequence of BB4 from sequencing: confirmed [X]
-> Sequence of BB5 from sequencing: confirmed [ ]
-> Sequence of BB6 from sequencing: confirmed [ ]
Assembling Construct 2
Assembling Construct 3
- 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
- Sequencing
- Transform into HeLa cells
- Western Blot or Proteinchromatography
-> Check if PhiC31o is read off
Contents
Aim 1: DNA reproduction+PCR
PCR1: replication of the Tet-inducible CMV promotor [X]
PCR4a: replication human bak with mutagenesis (Pst1) [ ]
PCR4b: replication human bak with mutagenesis (Pst1) [ ]
PCR5: joining PCR of human bak [ ]
PCR6: replication of the SV40-polyadenylation site [X]
PCR9: replication of eGFP with attP in primer [ ]
PCR10: Replikation of PhiC31o [X]
Aim 2: Inserting PCR Products in pSB1C3 and verifying Sequence
Aim 3: Assembling Biobricks
Assembling Construct 1
Aim 4: Testing products
Construct 1
Construct 2
Construct 3
Aim 5: Testing system
