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Table 2 PCR conditions for each pathogens investigated

From: Comprehensive study of ticks and tick-borne diseases in dogs in Nepal: molecular identification, risk analysis and hematological alterations

Pathogens

Hot Start condition

Amplification Cycles

Final extension at 72 °C

Reference

Conditions

No.

Babesia

First Round

94 °C for 3 min, 58 °C for 1 min, 72 °C for 2 min

94 °C for 30 s, 58 °C for 20 s, 72 °C for 30 s

45

7 min

(55)

Second Round

94 °C for 3 min, 58 °C for 1 min, 72 °C for 2 min

94 °C for 30 s, 62 °C for 20 s, 72 °C for 30 s

H. canis

95 °C for 5 min

95 °C for 30s, 57 °C for 30s, 72 °C for 90s,

40

5 min

(56)

E. canis

94 °C for 3 min

94 °C for 30 s, 62 °C for 30 s, 72 °C for 30 s

2

3 min

(57)

94 °C for 30 s, 60 °C for 30 s, 72 °C for 30 s

2

94 °C for 30 s, 58 °C for 30 s, 72 °C for 30 s

2

94 °C for 30 s, 56 °C for 30 s 72 °C for 30 s

2

94 °C for 30 s, 54 °C for 30 s 72 °C for 30 s

2

94 °C for 30 s, 52 °C for 30, 72 °C for 30 s

39

Borrelia

94 °C for 2 min

94 °C for 30s, 58 °C for 1 min, 72 °C for 1 min

40

1 min

(58)

A. platys

95 °C for 5 min

95 °C for 30s, 55 °C for 30s, 72 °C for 90

40

5 min

(54)