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Table 2 Thermal cycling parameters used in SWPOP-PCR method

From: Stepwise partially overlapping primer-based PCR for genome walking

Round of PCR Stage Thermal condition Cycle number
Primary   94 °C 2 min 1
1 94 °C 30 s, 65 °C 1 min, 72 °C 2 min 5
2 94 °C 30 s, 25 °C 1 min, 72 °C 2 min 1
3 94 °C 30 s, 65 °C 1 min, 72 °C 2 min 25
  72 °C 5 min 1
1 μL of the product was directly used as plate in the secondary round of PCR
Secondary   94 °C 2 min 1
1 94 °C 30 s, 65 °C 1 min, 72 °C 2 min 5
2 94 °C 30 s, 50 °C 1 min, 72 °C 2 min 1
3 94 °C 30 s, 65 °C 1 min, 72 °C 2 min 25
  72 °C 5 min 1
1 μL of the product was directly used as plate in the tertiary round of PCR
Tertiary Reaction profile of tertiary PCR is identical to that of secondary PCR