Figure 3.

Optimizing the PCR conditions. (a) Neither extending the denaturation times (dark red squares) nor adding 2M betaine (black triangles) is sufficient to recover extremely GC-rich DNA fragments by PCR with Phusion HF. (b) Combining long denaturation and 2M betaine is effective for the high-GC fraction (black triangles) but the profile is not as even over the entire GC spectrum as after PCR with AccuPrime Taq HiFi (blue diamonds) using extended denaturation times and a lower temperature (65°C) for primer annealing and extension.