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Table 2 Thermal cycling parameters for the DAR-PCR method

From: DAR-PCR: a new tool for efficient retrieval of unknown flanking genomic DNA

Round of PCR

Stage

Thermal conditions

Cycle number

Primary

 

94 °C, 2 min

 

1

94 °C 30 s, 60 °C 30 s, 72 °C 2 min

5

2

94 °C 30 s, 25 °C 30 s, 72 °C 2 min

1

3

94 °C 30 s, 55 °C 30 s, 72 °C 2 min

15

4

94 °C 30 s, 65 °C 30 s, 72 °C 2 min

25

  

72 °C 5 min

 

1 µL of the product was directly used as template for secondary PCR

Secondary

 

94 °C, 2 min

 

1

94 °C 30 s, 60 °C 30 s, 72 °C 2 min

35

 

72 °C 5 min