Sean Lauber:PCR: Difference between revisions
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(New page: For each reaction: 18.15 μL nuclease free water 2.5 μL 10X PCR buffer 0.5 μL 10 μM dNTPs 0.75 μL 50 mM MgCl2 1 μL 25 μM primer A 1 μL 25 μM primer B 1 μL ...) |
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Revision as of 08:18, 11 July 2012
For each reaction:
18.15 μL nuclease free water 2.5 μL 10X PCR buffer 0.5 μL 10 μM dNTPs 0.75 μL 50 mM MgCl2 1 μL 25 μM primer A 1 μL 25 μM primer B 1 μL template (about 100 ng) 0.1 μL Taq (platinum taq is good!)
25 μL Total
Place in a thermocycler and program it as follows:
94°C 3 min
94°C 0.5 min| x°C 0.5 min| x35 72°C 1 min |
72°C 1 min
x: annealing temperature, this varies (search for annealing temp calculator to estimate from primer sequence)
