Table 1.

Primer set used for RT-PCR amplification of the eight vRNAs of B/Yamanashi/166/98

Gene	Forward primer	Reverse primer
PB1	Bm-PB1b-1:	Bm-PB1b-1200R:
[1A]	TATTCGTCTCAGGGAGCAGAAGCGGAGCCTTTAAGATG	TATTCGTCTCGATGCCGTTCCTTCTTCATTGAAGAATGG
PB1	Bm-PB1b-1220:	Bm-PB1b-2369R:
[1B]	TATTCGTCTCGGCATCTTTGTCGCCTGGGATGATGATG	ATATCGTCTCGTATTAGTAGAAACACGAGCCTT
PB2	Bm-PB2b-1:	Bm-PB2b-1145R:
[2A]	TATTCGTCTCAGGGAGCAGAAGCGGAGCGTTTTCAAGATG	TATTCGTCTCTCTCATTTTGCTCTTTTTTAATATTCCCC
PB2	Bm-PB2b-1142:	Bm-PB2b-2396R
[2B]	TATTCGTCTCATGAGAATGGAAAAACTACTAATAAATTCAGC	ATATCGTCTCGTATTAGTAGAAACACGAGCATT
PA	Bm-PAb-1:	Bm-PAb-1261R:
[3A]	TATTCGTCTCAGGGAGCAGAAGCGGTGCGTTTGA	TATTCGTCTCCCAGGGCCCTTTTACTTGTCAGAGTGC
PA	Bm-PAb-1283:	Bm-PAb-2308R:
[3B]	TATTCGTCTCTCCTGGATCTACCAGAAATAGGGCCAGAC	ATATCGTCTCGTATTAGTAGAAACACGTGCATT
HA	MDV-B 5′ BsmBI-HA:	MDV-B 3′ BsmBI-HA:
	TATTCGTCTCAGGGAGCAGAAGCAGAGCATTTTCTAATATC	ATATCGTCTCGTATTAGTAGTAACAAGAGCATTTTTC
NP	MDV-B 5′ BsmBI-NP:	MDV-B 3′ BsmBI-NP:
	TATTCGTCTCAGGGAGCAGAAGCACAGCATTTTCTTGTG	ATATCGTCTCGTATTAGTAGAAACAACAGCATTTTTTAC
NA	Bm-NAb-1:	Bm-NAb-1557R:
	TATTCGTCTCAGGGAGCAGAAGCAGAGCA	ATATCGTCTCGTATTAGTAGTAACAAGAGCATTTT
M	MDV-B 5′ BsmBI-M:	MDV-B 3′ BsmBI-M:
	TATTCGTCTCAGGGAGCAGAAGCACGCACTTTCTTAAAATG	ATATCGTCTCGTATTAGTAGAAACAACGCACTTTTTCCAG
NS	MDV-B 5′ BsmBI-NS:	MDV-B 3′ BsmBI-NS:
	TATTCGTCTCAGGGAGCAGAAGCAGAGGATTTGTTTAGTC	ATATCGTCTCGTATTAGTAGTAACAAGAGGATTTTTAT

The sequences complementary to the influenza sequences are shown in bold. The 5′-ends have recognition sequences for the restriction endonuclease BsmBI (Bm, CGTCTCN1/N5). The design of the primers for PB1, PB2, and PA allowed the amplification of two fragments (1A, 1B, 2A, 2B, 3A, 3B).