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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