Jones Lab:Protocols:PCR: Difference between revisions
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(New page: <h3>PCR</h3> Add the following to a PCR tube (Master mix if necessary) #10 μL H<sub>2</sub>O #2 μL 10x PCR buffer #2 μL 2.5mM dNTPs (each) #2 μL 20mM MgCl<sub>2</sub> #1 μL Forward ...) |
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Latest revision as of 01:01, 31 October 2012
PCR
Add the following to a PCR tube (Master mix if necessary)
- 10 μL H2O
- 2 μL 10x PCR buffer
- 2 μL 2.5mM dNTPs (each)
- 2 μL 20mM MgCl2
- 1 μL Forward primer (10μM)
- 1 μL Reverse primer (10μM)
- 1 μL taq polymerase
- 1 μL DNA
Mix gently, then setup PCR cycles
- 94°C for 2 mins
- 94°C for 30 secs
- 58°C for 15 secs
- 72°C for 1 min per Kb of amplicon
- Go to #2 35 times
- 72°C for 5 mins
- 10°C hold