TABLE 3.

Primers used for PCR and RT-PCR

Target region	Forward primer sequencea (5′-3′)	Tm (°C)b	Reverse primer sequencea (5′-3′)	Tm (°C)	Amplicon length (bp)
1 (NS2a)	WNV_PCR_F5, GCCAACTACCGCAACACAGYTGGGCCTTMTGGTCGTGTTC	81-83	WNV_PCR_R7A, CCAACTACCGCAACCACTTTGATGAGGCTTCCAACTCCAACCAT	81	540-580
	WNV_PCR_F6, GCCAACTACCGCAACACTGGCCACCCAGGAGGTCCTTC	83	WNV_PCR_R7B, CCAACTACCGCAACCACCCTGATCAARCTGCCTATTCCRACCAT	81-83
			WNV_PCR_R8A, CCAACTACCGCAACCAATGAGGCAAGCTCCTTTCTTTTTTGC	79
			WNV_PCR_R8B, CCAACTACCGCAACCAARCAGACTKGCTCCTTTCTTYTTTGC	78-81
2 (NS5)	WNV_PCR_F9A, GCCAACTACCGCAACACGATGTGGAAGAGGCGGYTGGTGTTA	82-83	WNV_PCR_R11A, CCAACTACCGCAACCACCCAGAAGCACCTGGCTWGTCAT	81	460-510
	WNV_PCR_F9B, GCCAACTACCGCAACACGGTGTGGTAGAGGCGGCTGGTGCTA	85	WNV_PCR_R11B, CCAACTACCGCAACCACCTARGAGYACCTGGCTGGTCAT	80-82
	WNV_PCR_F10A, GCCAACTACCGCAACACAGAAGAGGGTRCAGGAAGTGAAAGGGTACAC	83-84	WNV_PCR_R12A, CCAACTACCGCAACCAGGTCCCTTCCAKGTYTTCTTYTCCAT	79-82
	WNV_PCR_F10B, GCCAACTACCGCAACACAAAAAAGAGTYCAAGAAGTCAGAGGGTACAC	81	WNV_PCR_R12B, CCAACTACCGCAACCAGGTCCCTTCCAGGTCCTYTTYTCCAT	81-83
3 (NS5)	WNV_PCR_F13A, GCCAACTACCGCAACACGCGAACCACCCCTACAGGACCTGGAAC	85	WNV_PCR_R15A, CCAACTACCGCAACCAGGYTTTTTCTCYCTCTTTCCCATC	78-80	420-560
	WNV_PCR_F13B, GCCAACTACCGCAACACGAGAACCACCCATATAGAACCTGGAAC	82	WNV_PCR_R15B, CCAACTACCGCAACCAGGTTTCTTYTCTCTCTTSCCCATC	79-80
	WNV_PCR_F14A, GCCAACTACCGCAACACCAATGTGACCACGATGGCCATGAC	82	WNV_PCR_R16A, CCAACTACCGCAACCACAGAAAGCGAGCTCCKAGCCAC	81-82
	WNV_PCR_F14B, GCCAACTACCGCAACACGAATGTYACCACMATGGCCATGAC	80-82	WNV_PCR_R16B, CCAACTACCGCAACCACAGGAAGCGRGCYCCCAGCCAC	83-85

^aThe universal tail sequences appended to the 5′ ends of all primers are underlined.

^bT_m, melting temperature.