Muratore:Protocols/PCR/Basic
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Tube | sterile H2O | Pfu Buffer | Template | 5'primer | 3'primer | dNTPs | Pfu Turbo | wax |
---|---|---|---|---|---|---|---|---|
(10X) | (10 μg/mL) | (4 μM) | (4 μM) | (10 mM ea) | (2.5 U/μL) | |||
Experimental | 72 μL | 10 μL | 5 μL | 5 μL | 5 μL | 2 μL | 1 μL | 50 μL |
(-) Control | 73 μL | 10 μL | 5 μL | 5 μL | 5 μL | 2 μL | 0 μL | 50 μL |
- Add each from left to right. Mix and spin before adding Pfu Turbo. Gentle mixing and spin after Pfu Turbo. Don't mix in wax.
- 5' @ 95°C
- 30" @ 95°C
- 2' @ Tm-5°C
- 2' @ 78°C
- repeat steps 2-4 x 29 more times
- 5' @ 78°C