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Table 1 PCR reaction components

From: A restriction-free method for gene reconstitution using two single-primer PCRs in parallel to generate compatible cohesive ends

  Reaction 1 Reaction 2a Reaction 2b
Template DNA ~50 ng ~500 ng ~500 ng
Forward primer (100 μM) 0.25 μL 0.25 μL  
Reverse primer (100 μM) 0.25 μL   0.25 μL
Phusion GC Buffer (5×) 10 μL 10 μL 10 μL
dNTPs (10 mM) 1 μL 1 μL 1 μL
DMSO (100%) 1.5 μL 1.5 μL 1.5 μL
Phusion High Fidelity DNA
Polymerase
1 μL 1 μL 1 μL
Add water to 50 μL 50 μL 50 μL