IGEM:UNAM/2009/Notebook/Modeling logbook Claudia/2010/10/02

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Ligation Procedure:Lumazine/LuxY and P0451 with promoter J23102.

I'm working in Step 7 of the Ligation Procedure: Lumazine, LuxY and P0451 with promoter J23102.

  • 7.Colony PCR to confirm ligations:

Analyze the colonies with Colony PCR to confirm that they contain the correct ligation.

The primers that I am using are:

Forward (5'->3'): Preffix primer.

Reverse:(5'->3'): Suffix primer.

These primers would amplify Lumazine, LuxY and P0451 ligated to each promoter, if the ligations were correctly done.

Results:Ligations Lumazine, LuxY and P0451 with promoter J23102.Procedure Step 7

  • Colonies selected to do the colony PCR reactions:

P0451 + J23102 ligation from colonies 1,2 and 31.

Colony PCRs:Ligations Lumazine with promoter J23102..Lane1:Ladder. Lanes 11,12 and 13: PCR amplified product P0451 + J23102 colonies 1, 5 and 8,respectively. The other lanes are samples from other experiments.
Colony PCRs:Ligations Lumazine with promoter J23102..Lane1:Ladder. Lanes 11,12 and 13: PCR amplified product P0451 + J23102 colonies 1, 5 and 8,respectively. The other lanes are samples from other experiments.


LuxY + J23102 ligation from colonies 2, 1 and 4.

Colony PCRs:Ligations LuxY with promoter J23102..Lane1:Ladder. Lane 14 and 15: PCR amplified product LuxY (Mr.gene) + J23102 colonies 2, 1 and 4 respectively. The other lanes are samples from other experiments.
Colony PCRs:Ligations LuxY with promoter J23102..Lane1:Ladder. Lane 14 and 15: PCR amplified product LuxY (Mr.gene) + J23102 colonies 2, 1 and 4 respectively. The other lanes are samples from other experiments.
Colony PCRs:Ligations LuxY with promoter J23102..Lane1:Ladder. Lane 2: PCR amplified product LuxY (Mr.gene) + J23102 colony 4.The other lanes are samples from other experiments.
Colony PCRs:Ligations LuxY with promoter J23102..Lane1:Ladder. Lane 2: PCR amplified product LuxY (Mr.gene) + J23102 colony 4.The other lanes are samples from other experiments.


Lumazine + J23102 ligation from colony 1,5 and 8.

Colony PCRs:Ligations Lumazine with promoter J23102..Lane1:Ladder. Lanes 8,9 and 10: PCR amplified product Lumazine (Mr.gene) + J23102 colonies 1, 5 and 8,respectively. The other lanes are samples from other experiments.
Colony PCRs:Ligations Lumazine with promoter J23102..Lane1:Ladder. Lanes 8,9 and 10: PCR amplified product Lumazine (Mr.gene) + J23102 colonies 1, 5 and 8,respectively. The other lanes are samples from other experiments.

Lumazine + J23102 ligation from colony 2,3,4,6 and 7.

Colony PCRs:Ligations Lumazine with promoter J23102..Lane1:Ladder. Lanes 2,3,4,5 and 6: PCR amplified product Lumazine (Mr.gene) + J23102 colonies 2,3,4,6 and 7,respectively. The other lanes are samples from other experiments.
Colony PCRs:Ligations Lumazine with promoter J23102..Lane1:Ladder. Lanes 2,3,4,5 and 6: PCR amplified product Lumazine (Mr.gene) + J23102 colonies 2,3,4,6 and 7,respectively. The other lanes are samples from other experiments.

Ligation Procedure: cI inverter(P0451 + K098991) with plasmid pSB1C3. Step 7

I'm working in Step 7 of the Ligation Procedure: cI inverter construction with pSB1C3 backbone.

  • 7.Colony PCR to confirm ligations:

Analyze the colonies with Colony PCR to confirm that they contain the correct ligation.

The primers that I am using are:

Forward (5'->3'): Preffix primer.

Reverse:(5'->3'): Suffix primer.

These primers would amplify the cI inverter (Parts P0451 + K098991) inside plasmid pSB1C3, if the ligation was correctly done.

Results: Ligation Procedure: cI inverter(P0451 + K098991) with plasmid pSB1C3. Step 7

  • Colonies selected to do the colony PCR reactions:

cI inverter(P0451 + K098991) with plasmid pSB1C3 ligation from colonies 2,3,4,5 and 6.

Colony PCRs:Ligation cI inverter( Parts P0451 + K098991) with plasmid pSB1C3..Lane1:Ladder. Lanes 3,4,5,6 and 7: PCR amplified product P0451 + J23102 colonies 2,3,4,5 and 6;respectively. The other lanes are samples from other experiments.
Colony PCRs:Ligation cI inverter( Parts P0451 + K098991) with plasmid pSB1C3..Lane1:Ladder. Lanes 3,4,5,6 and 7: PCR amplified product P0451 + J23102 colonies 2,3,4,5 and 6;respectively. The other lanes are samples from other experiments.



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