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Table 2 Primer characteristics of cytokine and reference genes for bovine, caprine and ovine gene expression

From: Design and evaluation of a unique SYBR Green real-time RT-PCR assay for quantification of five major cytokines in cattle, sheep and goats

Target gene Primer sequence (5′-3′) Optimal primer concentration (nM) Amplicon size (bp)
IL-4 F-CAGCATGGAGCTGCCT 300 177
R-ACAGAACAGGTCTTGCTTGC 300
IL-10 F-CTTTAAGGGTTACCTGGGTTGC 300 239
R-CTCACTCATGGCTTTGTAGACAC 300
IL-12B F-CAGCAGAGGCTCCTCTGAC 600 237
R-GTCTGGTTTGATGATGTCCCTG 600
INF-γ F-CAGAGCCAAATTGTCTCCTTC 300 167
R-ATCCACCGGAATTTGAATCAG 300
TNF-α F-CCAGAGGGAAGAGCAGTCC 300 111
R-GGCTACAACGTGGGCTACC 300
GAPDH F–ATCTCGCTCCTGGAAGATG 600 227
R-TCGGAGTGAACGGATTCG 300
H3F3A F-GAGGTCTCTATACCATGGCTC 300 150
R-GTACCAGGCCTGTAACGATG 300
ACTB F-TGGGCATGGAATCCTG 600 194
R-GGCGCGATGATCTTGAT 600
PPIA F-TGACTTCACACGCCATAAT 300 180
R-CTTGCCATCCAACCACTC 600
YWHAZ F-GAAAGGGATTGTGGACCAG 300 183
R-GGCTTCATCAAATGCTGTCT 300
  1. Abbreviations. F: forward; R: reverse; bp: base pair; IL, interleukin; IL12B, interleukin p40; TNF-α tumor necrosis factor-alpha; IFN-γ interferon-gamma; ACTB, beta-actin; GAPDH, Glyceraldehyde 3-phosphate dehydrogenase; H3F3A, H3-histone family 3A; PPIA, peptidylprolyl isomerase A; YWHAZ, tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, zeta polypeptide;
  2. Amplicon sizes were determinate in Bos taurus. Amplicon sizes in Capra hircus and Ovis aries were similar.