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Team:Calgary/3 July 2009
From 2009.igem.org
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Purpose: to verify the presence of LuxPQ into psB1AC3 by colony PCR; to isolate plasmid; to send isolated plasmid for sequencing. A pTaq colony PCR was set up using the following sets of forward and reverse primers: LuxPQ and BBK CP. The following conditions were used for both sets of primers: 94ºC for 6 minutes; 36X (94ºC for 30 seconds; 55ºC for 45 seconds; 72ºC for 4 minutes); 72ºC for 10 minutes; held at 4ºC. The products were then run on a 0.8% agarose gel (see figures below). We can see that colonies 1, 6 and 7 all have bands of the expected size (~3.9kb) for both primers, indicating a successful transformation of LuxPQ into psB1AC3. Plasmid was isolated from these colonies using the GenElute Plasma MinPrep Kit. Colonies 6 and 7 were then sent to the University of Calgary’s DNA sequencing laboratory with BBK-CP-F/R primers. | Purpose: to verify the presence of LuxPQ into psB1AC3 by colony PCR; to isolate plasmid; to send isolated plasmid for sequencing. A pTaq colony PCR was set up using the following sets of forward and reverse primers: LuxPQ and BBK CP. The following conditions were used for both sets of primers: 94ºC for 6 minutes; 36X (94ºC for 30 seconds; 55ºC for 45 seconds; 72ºC for 4 minutes); 72ºC for 10 minutes; held at 4ºC. The products were then run on a 0.8% agarose gel (see figures below). We can see that colonies 1, 6 and 7 all have bands of the expected size (~3.9kb) for both primers, indicating a successful transformation of LuxPQ into psB1AC3. Plasmid was isolated from these colonies using the GenElute Plasma MinPrep Kit. Colonies 6 and 7 were then sent to the University of Calgary’s DNA sequencing laboratory with BBK-CP-F/R primers. | ||
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[[Image:2009.07.02.BBKluxPQ_ColPCR_PQPrimers.png|700px]] | [[Image:2009.07.02.BBKluxPQ_ColPCR_PQPrimers.png|700px]] | ||
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[[Image:2009.07.02.BBKluxPQ_ColPCR_BBKCPPrimers.png|700px]] | [[Image:2009.07.02.BBKluxPQ_ColPCR_BBKCPPrimers.png|700px]] | ||
Revision as of 14:26, 9 September 2009
UNIVERSITY OF CALGARY
