TABLE 1.
Oligonucleotide primer and probe sequences for real-time PCR amplificationa
| Target | Oligonucleotide sequence (5′→3′)b | Concn (nM) | Product size (bp) |
|---|---|---|---|
| β2M | AGACACCCACCAGAAGATGG, F | 600 | 98 |
| TCCCCATTCTTCAGCAAATC, R | 600 | ||
| FAM-TGGGTTCCATCCACCCCAGA-BHQ, P | 300 | ||
| IFN-γ | GCGCAAAGCCATAAATGAAC, F | 900 | 98 |
| CTCAGAAAGCGGAAGAGAAG, R | 900 | ||
| HEX-CAAAGTGATGAATGACCTGTGCCA-BHQ, P | 300 |
a
Primers and probes were designed to detect β2M and IFN-γ of multiple species, including elk, cattle, sheep, goat, and bison, for which sequence information was available from EMBL/GenBank.
b
Probes contained either the 6-carboxyfluorescein (FAM) or hexachloro-6-carboxyfluorescein (HEX) reporter dye covalently attached at the 5′ end and the BHQ molecule covalently attached at the 3′ end. F, forward primer; R, reverse primer; P, probe.