Team:University of Lethbridge/Notebook/Project3July
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+ | [[Team:University_of_Lethbridge/Notebook|Back to The University of Lethbridge Main Notebook]] | ||
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===July 27, 2008=== | ===July 27, 2008=== | ||
====Nathan Phillips==== | ====Nathan Phillips==== | ||
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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 | ||
Line 19: | Line 21: | ||
3. Final extension @ 72C for 10 mins (1 cycle) | 3. Final extension @ 72C for 10 mins (1 cycle) | ||
4. Hold at 4C | 4. Hold at 4C | ||
+ | |||
+ | |||
+ | ===July 29, 2008=== | ||
+ | ====Nathan Puhl, Andrew, Alix==== | ||
+ | Objective: Optimize conditions for TIR PCR. | ||
+ | |||
+ | Set up PCR reaction for 5 reactions. | ||
+ | |||
+ | PCR cycle conditions: | ||
+ | 1. Initial denaturation @ 98 C for 30 sec (1 cycle) | ||
+ | 2a. Denaturation @ 98 C for 10 sec | ||
+ | 2b. Annealing @ 42 C for 30 sec | ||
+ | 2c. Extension @ 72 C for 15 sec (Repeat from step 2, 29 times) | ||
+ | 3. Final extension @ 72 C for 7 mins (1 cycle) | ||
+ | 4. Hold at 4 C |
Latest revision as of 16:37, 25 August 2008
Back to The University of Lethbridge Main Notebook
Contents |
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
July 29, 2008
Nathan Puhl, Andrew, Alix
Objective: Optimize conditions for TIR PCR.
Set up PCR reaction for 5 reactions.
PCR cycle conditions:
1. Initial denaturation @ 98 C for 30 sec (1 cycle) 2a. Denaturation @ 98 C for 10 sec 2b. Annealing @ 42 C for 30 sec 2c. Extension @ 72 C for 15 sec (Repeat from step 2, 29 times) 3. Final extension @ 72 C for 7 mins (1 cycle) 4. Hold at 4 C