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From 2012.igem.org

-> 20 µl / rxn

• 2 µl 10x Buffer
• 0.4 µl dNTPs (10 mM each)
• 0.8 µl Primer (10 µM) (2x or as premix)
• 0.4 µl Template (ca. 20-100 ng)
• 0.4 µl Pfu-Turbo
• ad 20 µl ddH2O

Program: -> 1 min 96°C

• 20 s 96°C
• 20 s 58°C
• 1.5 min/kb 72°C
• 15 Cycles

-> 5 min 72°C -> Store @ 4-8°C

After PCR add 1 µl DpnI directly into PCR tube. Incubate reaction 1-2 h at 37°C. Transform 5 µl into 50 µl competent cells.