IGEM:Arizona State/2014/Notebook/asu igem 2014/2014/08/29

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Entry title accD SOEing Used a Clean and Concentrate kit on both the Forward and Back megaprimers. Total volume = 20 μL Set up another PCR reaction to make final accD part SOEing PCR reaction 2 Reagent 2x GoTaq Master Mix 25 Forward Megaprimer (from PCR reactions on 8/27 ) 5.0 Reverse Megaprimer (from PCR reactions on 8/27 ) 5.0 H2O 15 Total 50.0 Initial denature step of 95 °C for 2 min PCR cycle: 95 °C for 30 sec -> 45 °C for 45 sec -> 73 °C for 80 sec Final annealing step at 73 °C for 5 min