Table 1.

Primers used for amplification and sequencing of 16S rDNA-, g19-, gp36-, and p28-specific genes for detection of Ehrlichia canis in blood samples from dogs

Target	Primers	Size (bp)	Reagents	Thermocycler	Reference
16S rDNA	F-5′TATAGCCTCTGGCTATAGGAAATTGTTA′3 R-5′ACCATTTCTAATGGCTATTCCGTACTA′3 5′6-FAMTGGCAGACGGGTGAGTAATGGTAGG-TAMRA-3′ (probe)	93 bp	TaqMan Universal Master Mix (ThermoFisher Scientific, Inc., Waltham, MA, USA), 1 × Primer, 0.3 μM (each) DNA, 3 μl Fv, 12 μl	50 °C, 2 min 95 °C, 10 min 45 cycles, 95 °C, 15 s; 60 °C, 1 min;	Baneth et al. 2008
gp19	F 5′-ATTAGTGTTGTGGTTATGCAA-3′ R 5′-TACGCTTGCTGAATATCATGA-3′	414 bp	Buffer (Tris-HCl 200 mM, pH 8.4, KCl 500 mM),1 × MgCl2, 2.5 mM dNTPs, 0.2 mM Primers, 0.6 μM (each) Taq, 0.125 U DNA, 5 μl Fv, 25 μl	94 °C, 3 min 35 cycles, 94 °C, 1 min; 55 °C, 1 min; 72 °C, 1.5 min; 72 °C, 5 min	Chen et al. 2010
p28	F 5′-ATGAATTGCAAAAAAATTCTTATA-3′ R 5′-TTAGAAGTTAAATCTTCCTCC -3′	843 bp	Buffer (Tris-HCl 200 mM, pH 8.4, KCl 500 mM), 1 × MgCl2, 2.5 mM dNTPs, 0.2 mM Primers, 0.6 μM (each) Taq, 1.2 U DNA, 5 μl Fv, 25 μl	95 °C, 5 min 30 cycles, 95 °C, 30 s; 55 °C, 1 min; 72 °C, 2 min; 72 °C, 5 min.	Nakaghi et al. 2010
gp36	F 5′-GTATGTTTCTTTTATATCATGGC-3′ R 5′-GGTTATATTTCAGTTATCAGAAG-3′	840 bp	Buffer (Tris-HCl 200 mM, pH 8.4, KCl 500 mM), 1 × MgCl2, 2.5 mM dNTPs, 0.2 mM Primers, 0.6 μM (each) Taq: 0.125 U DNA, 5 μl Fv, 25 μl	94 °C, 3 min 35 cycles, 94 °C, 1 min; 55 °C, 1 min; 72 °C, 1.5 min; 72 °C, 5 min	Chen et al. 2010

F, forward; R, reverse; bp, base pairs; min, minutes; sec, seconds; Fv, final volume; Taq polymerase and dNTPS-Invitrogen® (Thermo Fisher Scientific)