LovTAP fused to promoters and Lumazine:Ligations to backbone pSB3K3 and pSB1C3
I'm working in Step 7 of the Repeated experiment:Working on LovTAP fused to promoters and Lumazine:Ligations to backbone pSB3K3 and pSB1C3.
- 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 LovTAP fused to promoters and Lumazine, if the ligations were correctly done.
Results:LovTAP fused to promoters and Lumazine:Ligations to backbones pSB3K3 and pSB1C3.Procedure Step 7
- Colonies selected to do the colony PCR reactions:
Ligations to plasmid pSB3K3
LovTAP + J23117 isolated from colonies 2, 3 and 4.
LovTAP + J23105 isolated from colonies 1, 2 and 3.
LovTAP + J23114 isolated from colonies 9,13 and 14.
LovTAP + J23102 isolated from colonies 1,11 and 13.
Ligations to plasmid pSB1C3
LovTAP + J23117 isolated from colonies 1,6 and 7.
LovTAP + J23105 isolated from colonies 1, 12 and 13.
LovTAP + J23114 isolated from colonies 1,2 and 3.
LovTAP + J23102 isolated from colonies 1,2 and 8.
Lumazine (Mr.Gene) colonies 1 and 2.
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