TABLE 5.
Primers used for BUNV S-segment qRT-PCRsa
| Reaction | Target and primer or probe | Oligonucleotide sequence (5′ → 3′) | Position on RNA segment |
|---|---|---|---|
| RT | gRNA | GGCCGTCATGGTGGCGAATCTTGCTATTGTTGAAAATCGCTGTGCTA | 911–938 |
| cRNA | GCTAGCTTCAGCTAGGCATCCAATATAATGTTGATTTAGCCC | 903–924 | |
| mRNA | CCAGATCGTTCGAGTCGTCATCCCTGCTTACATGTTGATTC | 774–796 | |
| qPCR | gRNA forward primer | CGACATCATGAAATTCCAA | 849–867 |
| gRNA probe | FAM-ATCCAACAGAAGGTCATTAAAGGCTC-BHQ1 | 881–906 | |
| gRNA reverse primer | GGCCGTCATGGTGGCGAAT | ||
| cRNA forward primer | ACAAAATAACAGCTGCTTGG | 856–875 | |
| cRNA probe | FAM-TTTAGCCCGCTGTCTTTCTGTCCC-BHQ1 | 887–910 | |
| cRNA reverse primer | GCTAGCTTCAGCTAGGCATC | ||
| mRNA forward primer | CAGTTGTCTCTAGCTTAGGTTGG | 702–724 | |
| mRNA probe | FAM-TCCCTGGCAGCTGCACTAACATT-BHQ1 | 734–756 | |
| mRNA reverse primer | CCAGATCGTTCGAGTCGT |
a
Based on the S-segment sequence under GenBank accession number NC_001927.1. Italic sequences show the nonviral tags.