Team:University of Lethbridge/Notebook/Project3July

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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 PCR machine 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