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

UNAM-Genomics-Mexico team

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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. 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 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. 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. 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.