Table 2.
Oligonucleotides used for cloning of the supporting plasmids, construction of pFLC-LS1-1eGFP, and confirmation of virus recovery
Primers | Sequences (5′ → 3′)a,b | Restriction site | Nucleotide Positionc |
---|---|---|---|
Supporting plasmid primers | |||
Nfor | GGAATTC GCCACCATGTCTTCCGTATTTGATG | EcoRI | 116–140 |
Nrev | GCGGCCGCTCAATACCCCCAGTCGGTGT | NotI | 1572–1591 |
Pfor | TGAATTC GCCACCATGGCCACCTTTACAGATGC | EcoRI | 1886–1906 |
Prev | GCGGCCGCTTAGCCATTTAGAGCAAG | NotI | 3057–3074 |
Lfor | TACTAGT GCCACCATGGCGAGCTCCGGTCCTGAA | SpeI | 8380–8401 |
Lrev | GCGGCCGCTTAAGAGTCACAGTTACTGT | NotI | 14,976–14,995 |
pFLC-LS1-1eGFP construction | |||
NDV116_F | GCCAACATGTCTTCCGTATTTGATG | PciI | 116–140 |
NDV3211_R | CGATCATTCAGTGGGGCTGAGG | BbvCI | 3190–3211 |
Confirmation primers | |||
M + 4100 d | AGTGATGTGCTCGGACCTTC | N.A.e | 4100–4119 |
NDV4394_R | GAGACGCAGCTTATTTCTTAAAAGG | N.A. | 4370–4394 |
NDV14035_F | TCCAGGGTCCAATCAAAGCT | N.A. | 14,035–14,054 |
NDV15003_R | GATTTTCGTTAAGAGTCACAGTTACTG | N.A. | 14,977–15,003 |