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Team:University of Lethbridge/Notebook/Project3July
From 2008.igem.org
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| - | PCR cycle conditions (programmed into HJ's | + | PCR cycle conditions (programmed into HJ's thermocycler under "'''Nate'''": |
1. Initial denaturation @ 95C for 3 mins (1 cycle) | 1. Initial denaturation @ 95C for 3 mins (1 cycle) | ||
2a. Denaturation @ 95C for 30 sec | 2a. Denaturation @ 95C for 30 sec | ||
Revision as of 17:20, 29 July 2008
July 27, 2008
Nathan Phillips
Objective: Amplify TIR gene from E.coli total DNA from DH5a
PCR reaction setup:
-Enzyme = Phusion -Template = DH5a DNA -Buffer = HF Phusion buffer -Primers = IDT, designed & shipped July 16/08
PCR cycle conditions (programmed into HJ's thermocycler under "Nate":
1. Initial denaturation @ 95C for 3 mins (1 cycle) 2a. Denaturation @ 95C for 30 sec 2b. Annealing @ 49C 2c. Extension @ 72C for 30 sec (Repeat from step 2, 34 times) 3. Final extension @ 72C for 10 mins (1 cycle) 4. Hold at 4C
