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Team:Calgary/20 June 2009
From 2009.igem.org
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| - | Purpose: to verify presence of LuxPQ in psB1AK3. Colony 4 of luxPQ in psB1AK3 was used as a template and pTaq DNA polymerase was used. LuxPQ-F/R gene specific primers and BBK-CP-F/R primers (that anneal just outside the BBK prefix and suffix) were used. Conditions were as follows: 94ºC for 2 minutes; 36X (94ºC for 30 seconds; 55ºC for 45 seconds for BBK-CP-F/R primers (53ºC for 45 seconds for LuxPQ 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 (see below). | + | Purpose: to verify presence of LuxPQ in psB1AK3. Colony 4 of luxPQ in psB1AK3 was used as a template and pTaq DNA polymerase was used. LuxPQ-F/R gene specific primers and BBK-CP-F/R primers (that anneal just outside the BBK prefix and suffix) were used. Conditions were as follows: 94ºC for 2 minutes; 36X (94ºC for 30 seconds; 55ºC for 45 seconds for BBK-CP-F/R primers (53ºC for 45 seconds for LuxPQ 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 (see below). Again, we are looking for a band size of just under 4kb, which we do not see here, and we must thus restart from luxPQ in TOPO. |
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[[Image:2009.06.22.LuxPQ_BBKCP_BBK.png|700px]] | [[Image:2009.06.22.LuxPQ_BBKCP_BBK.png|700px]] | ||
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Revision as of 19:21, 2 September 2009
UNIVERSITY OF CALGARY
