Table 2.
Reverse primers1 used in 5'RACE for amplification of 5' non-coding region of different ISAV segments using mRNA transcripts as template
| ISAV RNA segment | Primer sequence |
|---|---|
| Segment 1 | ssp1: 5'-GCTCCAACTGCTGCTCTAC-3' |
| ssp2: 5'-CAGTACTCCTCGGTAGTTGG-3' | |
| Segment 2 | ssp1: 5'-GTGTTCAATCAGGTCTGACAT-3' |
| ssp2: 5'-CTCTGAAACCTGTTGCTACTAT-3' | |
| Segment 3 | ssp1: 5'-GTAACATCTGCCATGTCCAC-3' |
| ssp2: 5'-CTTCTGACAGGATTGCGCTG-3' | |
| Segment 4 | ssp1: 5'-CGTATGGCCCTCCTCTGTAA-3' |
| ssp2: 5'-CATGTGCTGATCGGTGTTGG-3' | |
| Segment 5 | ssp1: 5'-GTCTGGTACAGAATGGAACG-3' |
| ssp2: 5'-GTGCAACAGCTATCCAAGTC-3' | |
| Segment 6 | ssp1: 5'-GCGTCTGCTCGTCCAACAAGT-3' |
| ssp2: 5'-GCGTTGTCCAGTGTCATCGA-3' | |
| Segment 7 | ssp1: 5'-CAGACACTCCTCATAGTACC-3' |
| ssp2: 5'-GCATGTGTGACAGACCATC-3' | |
| Segment 8 | ssp1: 5'-CATCTGCATCCTGCTGTGTAG-3' |
| ssp2: 5'-CTTTCATCACACAGTAGAGAG-3' | |
1 The 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).