Muratore:Protocols/PCR/Basic

Basic PCR protocol

Basic PCR protocol

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.

1. 5' @ 95°C
2. 30" @ 95°C
3. 2' @ T_m-5°C
4. 2' @ 78°C
5. repeat steps 2-4 x 29 more times
6. 5' @ 78°C