|
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
|
- 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;
- Amplicon sizes were determinate in Bos taurus. Amplicon sizes in Capra hircus and Ovis aries were similar.
