Table 7.
Reverse primers1 used in 5'RACE for amplification of 5' non-coding region of different ISAV segments using vRNA as template
| ISAV RNA segment | Primer sequence |
|---|---|
| Segment 1 | ssp1: 5'-GTAGTGGCAAGTGCTAGATAC-3' |
| ssp2: 5'-GCCATGAGGTGCTGCATTG-3' | |
| Segment 2 | ssp1: 5'-GAAGCTGTGAACAGAGGACAT-3' |
| ssp2: 5'-GCTGCAGCATAGAGTTCGAT-3' | |
| Segment 3 | ssp1: 5'-GGTACAGATCCAACATCAAG-3' |
| ssp2: 5'-GCAAGTGAACGTGACTGACA-3' | |
| Segment 4 | ssp1: 5'-CTCTGTGCAAACAAAAGCAGA-3' |
| ssp2: 5'-TGGCAACATGACCCTGAGCT-3' | |
| Segment 5 | ssp1: 5'-GAGGAGGATTGCCTATATGAAC-3' |
| ssp2: 5'-CAAGTGGAAATCGGACTAATAG-3' | |
| Segment 6 | ssp1: 5'-GATGAGTACGTTGACACAC-3' |
| ssp2: 5'-CAGAGGCACTGACATGTCCA-3' | |
| Segment 7 | ssp1: 5'-CACTTGCCCAGACGAGTAC-3' |
| ssp2: 5'-CAGTGACAAGGACGAGATGC-3' | |
| Segment 8 | ssp1: 5'-GATCAGTCGAGCGACGATGA-3' |
| ssp2: 5'-GCATCCTGCTCGACAGAGAA-3' | |
1The forward primer for PCR in all cases (i.e., with either ssp1 or ssp2) was the Universal Amplification Primer (UAP) that comes with the 5'RACE kit (Invitrogen).