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Team:Calgary/25 June 2009
From 2009.igem.org
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Purpose: to verify the presence of LuxPQ in a BBK vector. Colonies 8, 9, 10, and 11 of LuxPQ in psB1AK3 were screened by pTaq colony PCR with the following sets of forward and reverse primers: LuxPQ, BBK and BBK CP. The following conditions were used: 94ºC for 6 minutes; 36X (94ºC for 30 seconds; 52ºC for 45 seconds for LuxPQ-F/R primers (or 50ºC for 45 seconds for BBK F/R primers, or 52ºC for 45 seconds for BBK CP F/R primers); 72ºC for 4 minutes); 72ºC for 10 minutes; held at 4ºC. The products were then run on a 0.8% agarose gel, along with a restriction digest of colonies 1-7 with NotI (see below). | Purpose: to verify the presence of LuxPQ in a BBK vector. Colonies 8, 9, 10, and 11 of LuxPQ in psB1AK3 were screened by pTaq colony PCR with the following sets of forward and reverse primers: LuxPQ, BBK and BBK CP. The following conditions were used: 94ºC for 6 minutes; 36X (94ºC for 30 seconds; 52ºC for 45 seconds for LuxPQ-F/R primers (or 50ºC for 45 seconds for BBK F/R primers, or 52ºC for 45 seconds for BBK CP F/R primers); 72ºC for 4 minutes); 72ºC for 10 minutes; held at 4ºC. The products were then run on a 0.8% agarose gel, along with a restriction digest of colonies 1-7 with NotI (see below). | ||
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[[Image:2009.06.24.LuxPQ_Col_PCR+BBK_Primer_Verif-1.png|700px]] | [[Image:2009.06.24.LuxPQ_Col_PCR+BBK_Primer_Verif-1.png|700px]] | ||
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Revision as of 16:15, 3 September 2009
UNIVERSITY OF CALGARY
