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

Table 4 Assay performances for reference genes for bovine, caprine and ovine gene expression

Target gene	Species	Tm (°C)	Intercept	Slope	E (%)	Inter-assay variability				LOD nc
Target gene	Species	Tm (°C)	Intercept	Slope	E (%)	Mean Cq	SD Cq	Mean nc	SD nc	LOD nc
*GAPDH*	cattle	84.5	2.02	−3.372	98	18.3	0.2	6.2 10⁴	0.7 10⁴	1
	goat		1.37	−3.276	102	19.6	0.2	1.1 10⁴	0.2 10⁴	1
	sheep		1.39	−3.283	102	24.9	0.1	285	12	2
*H3F3A*	cattle	85	0.27	−3.50	93	16.8	0.1	11.4 10⁴	0.5 10⁴	20
	goat		0.91	−3.33	100	16.4	0.1	14.2 10⁴	0.7 10⁴	11
	sheep		1.12	−3.34	99	16.5	0.1	15.4 10⁴	1.1 10⁴	14
*ACTB*	cattle	85.7	0.8	−3.32	100	15.4	0.4	19.4 10⁴	5.1 10⁴	8
	goat		1.70	−3.28	102	15.7	0.2	25.6 10⁴	3.8 10⁴	1
	sheep		1.59	−3.31	101	15.6	0.3	27.8 10⁴	0.5 10⁴	6
*PPIA*	cattle	82.3	1.27	−3.449	95	17.9	0.1	7.7 10⁴	0.5 10⁴	6
	goat		1.63	−3.309	100	17.5	0.1	8.3 10⁴	0.6 10⁴	3
	sheep		1.02	−3.409	96	17.4	0.1	8.2 10⁴	0.5 10⁴	8
*YWHAZ*	cattle	81.3	2.18	−3.417	96	18.7	0.1	7.2 10⁴	0.7 10⁴	9
	goat		0.38	−3.350	99	23.8	0.1	506	33	4
	sheep		0.04	−3.350	99	23.5	0.1	506	24	3

ISSN: 1746-6148