Table 1.

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

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

* product sizes and primer positions according to reference sequence AY046576