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Team:University of Lethbridge/Notebook/Protocols
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Contents |
PCR protocols
CheZ (Taq pol)
CheZ (Phusion pol)
Conditions for one reaction (25 uL):
-1x HF buffer (5 uL 5x buffer) -Forward primer (1.5 uL) -Reverse primer (1.5 uL) -dNTPs (0.5 uL) -Phusion (0.25 uL) -mQH2O (15.25 uL) -Template (1 uL)
Cycling protocol ("cheZ"):
1. Initial denaturation @ 98C for 4 mins (1 cycle) 2a. Denaturation @ 98 C for 30 sec (35 cycles for step #2) b. Annealing 47.0C for 30 seconds c. Extension @ 72C for 30 sec 3. Final extension @ 72C for 7 mins (1 cycle) 4. Hold at 4C
