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Team:Calgary/10 July 2009
From 2009.igem.org
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Purpose: To perform a colony PCR to verify the presence of the above constructs. Each Colony pTaq PCR reaction used the BBK CP F/R primers, each with the following similar conditions: 94ºC for 6 minutes; 36X (94ºC for 30 seconds; 55ºC for 45 seconds; 72ºC for 4 minutes for plasmid switch (or 6 min 10 sec for PQ-B-R-OU-B construct); 72ºC for 10 minutes; held at 4ºC. The products were then run on a 0.8% agarose gel (see below). For the plasmid switch, we can see that colonies 2, 5, 6, and 8 gave the desired size of LuxPQ (3.8kb), whereas in the signaling construct verification, we see desired sizes (~6.0kb) for colonies 11, 12 and 13. Next steps will be to isolate plasmid and proceed with further verification techniques on the colonies that appear to be what we want. | Purpose: To perform a colony PCR to verify the presence of the above constructs. Each Colony pTaq PCR reaction used the BBK CP F/R primers, each with the following similar conditions: 94ºC for 6 minutes; 36X (94ºC for 30 seconds; 55ºC for 45 seconds; 72ºC for 4 minutes for plasmid switch (or 6 min 10 sec for PQ-B-R-OU-B construct); 72ºC for 10 minutes; held at 4ºC. The products were then run on a 0.8% agarose gel (see below). For the plasmid switch, we can see that colonies 2, 5, 6, and 8 gave the desired size of LuxPQ (3.8kb), whereas in the signaling construct verification, we see desired sizes (~6.0kb) for colonies 11, 12 and 13. Next steps will be to isolate plasmid and proceed with further verification techniques on the colonies that appear to be what we want. | ||
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Revision as of 04:02, 10 September 2009
UNIVERSITY OF CALGARY
