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Table 1 Details of the nested and hemi-nested PCR protocols used to screen deer blood samples for the presence of Babesia spp.

From: Babesias of red deer (Cervus elaphus) in Ireland

PCR protocol

Primers (positions)*

 

Amplification protocol

Reference

I

1st PCR:

   

Product size*:

BTH-1F (365-385)

5'cct gag aaa cgg cta cca cat ct

94°C: 10 min,

[23]

561 bp

BTH-1R (1031-1050)

5' ttg cga cca tac tcc ccc ca

40 cycles: 95°C: 30 s, 68°C: 1 min, 72°C: 1 min

 
   

72°C: 10 min

 
 

nested PCR:

   
 

GF2 (466-487)

5' gtc ttg taa ttg gaa tga tgg

94°C: 10 min

[19]

 

GR2 (1006-1026)

5' cca aag act ttg att tct ctc

40 cycles: 95°C: 30 s, 60°C: 1 min, 72°C: 1 min

 
   

72°C: 10 min

 

II

1st PCR:

   

Product size*:

Babfor (959-977)

5'gac tag gga ttg gag gtc

94°C: 10 min

[24]

576 bp

Babrev (1589-1610)

5'gaa taa ttc acc gga tca ctc

35 cycles: 95°C: 1 min, 53°C: 1.5 min, 72°C: 1.5 min

 
   

72°C: 10 min

 
 

nested PCR

   
 

BT2-F (1035-1055)

5'gga gta tgg tcg caa gtc tg

94°C: 10 min

[23]

 

Babrev

 

35 cycles: 95°C: 1 min, 53°C: 1.5 min, 72°C: 1.5 min

 
   

72°C: 10 min

 
  1. * product sizes and primer positions according to reference sequence AY046576