Table 1. Characteristics of consensus primers (IL-4, IL-6, IL-10, IFNγ and TNFα) developed for Neophoca cinerea, Canis familiaris, Bos taurus, and Ovis aries in the study, and GAPDH (Peters et al., 2004) reference gene primers used in the study.
| Gene | GenBank ensemble sequences | Primers (5′->3′) (based on ** marked sequences) | Annealing/extension (°C, s) | Amplicon size (bp) | Conc (nM) | Tm (°C) |
|---|---|---|---|---|---|---|
| IL-4 | **AF083270.1_Canis familiaris | IL-4 F: TCACCTCCCAACTGATTCCAA | 60 °C, 30 | 135 | 200 | 82 |
| EU276069.1_Bos taurus | ||||||
| NM_001304911.1_Ailuropoda melanoleuca | ||||||
| KP792599.1_Neovison vison | ||||||
| XM_027606287.1_Zalophus californianus | IL-4 R: ACGAGTCGTTTCTCGCTGTG | 60 °C, 30 | ||||
| XM_006734683.1_Leptonychotes weddellii | ||||||
| AB020732.1_Tursiops truncatus | ||||||
| HM011505.1_Macropus eugenii | ||||||
| IL-6 | **L46802.1_Phoca vitulina | IL-6 F: CTGCTCCTGGTGATGGCTAC | 60 °C, 30 | 147 | 200 | 84.5 |
| U64794.1_Equus caballus | ||||||
| AF275796.1_Canis familiaris | ||||||
| EF368209.1_Mustela putorius furo | IL-6 R: TGCAGAGATTTTGCCGAGGA | 60 °C, 30 | ||||
| EF543744.1_Ailuropoda melanoleuca | ||||||
| L46803.1_Orcinus orca | ||||||
| IFNγ | **KJ888148.1_Neovison vison | IFNγ F: GTGAATGATCTGCAGGTCCA | 60 °C, 30 | 101 | 200 | 80 |
| NM_213948.1_Sus scrofa | ||||||
| NM_001003174.1_Canis familiaris | ||||||
| XM_026500086.1_Ursus arctos | ||||||
| XM_027593881.1_Zalophus californianus | IFNγ R: TGACTCCTTTTCCGCTTCCT | 60 °C, 30 | ||||
| XM_025883084.1_Callorhinus ursinus | ||||||
| XM_006740002.1_Leptonychotes weddellii | ||||||
| DQ118388.1_Phoca vitulina | ||||||
| IL-10 | **NM_001003077.1_Canis familiaris | IL-10 F: CTTTAAGAGTTACCTGGGTTGCC | 60 °C, 30 | 97 | 200 | 83.5 |
| DQ890062.1_Macaca mulatta | ||||||
| NM_001082490.1_Equus caballus | ||||||
| XM_002919274.3_Ailuropoda melanoleuca | ||||||
| XM_027613593.1_Zalophus californianus | IL-10 R: GATGTCTGGGTCGTGGTTCTC | 60 °C, 30 | ||||
| XM_004417581.2_Odobenus rosmarus | ||||||
| L46802.1_Phoca vitulina | ||||||
| AF026277.1_Trichosurus vulpecula | ||||||
| TNFα | **XM_027099307_Lagenorhynchus obliquidens | TNFα F: GAGCACTGAAAGCATGATCCG | 60 °C, 30 | 123 | 200 | 87 |
| D86587.1_Capra hircus | ||||||
| NM_001003244.4_Canis familiaris | ||||||
| EF368211.1_Mustela putorius furo | ||||||
| XM_002930032.3_Ailuropoda melanoleuca | TNFα R: GCGACCAGGAAGAAGGAGAA | 60 °C, 30 | ||||
| XM_027603858.1_Zalophus californianus | ||||||
| XM_025862490.1_Callorhinus ursinus | ||||||
| XM_006738478.1_Leptonychotes weddellii | ||||||
| GAPDH | Peters et al. (2004) | GAPDH F: TCAACGGATTTGGCCGTATTGG | 60 °C, 30 | 90 | 400 | 83.5 |
| GAPDH R: TGAAGGGGTCATTGATGGCG |
Notes:
Tm, melting temperature.
**
Marked sequences: IL-4 AF083270.1_Canis familiaris; IL-6 L46802.1_Phoca vitulina; IFNγ KJ888148.1_Neovison vison; IL-10 NM_001003077.1_Canis familiaris; TNFα XM_027099307_Lagenorhynchus obliquidens.