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 5 Absolute quantification of cytokine and reference gene expression in whole blood samples

Target gene	Species	Cq values		Copie number values
Target gene	Species	Mean Cq	SD Cq	Mean nc	SD nc
IL-4	cattle	28.7	0.3	24	5
	goat	28.5	0.2	31	3
	sheep	28.1	0.5	39	12
IL-10	cattle	27.2	0.6	156	54
	goat	28.2	0.5	48	17
	sheep	28.5	0.1	28	1
IL-12B	cattle	27.9	0.6	62	21
	goat	34.9	1.2	2	0.4
	sheep	33.3	0.5	2	0.7
INF-γ	cattle	27.4	0.4	106	28
	goat	27.7	0.3	103	21
	sheep	26.3	0.3	241	52
TNF-α	cattle	24.7	0.4	965	199
	goat	24.0	0.1	978	34
	sheep	24.4	0.1	728	34
*GAPDH*	cattle	18.0	0.3	7.6 10⁴	1.2 10⁴
	goat	19.1	0.1	1.6 10⁴	0.1 10⁴
	sheep	25.0	0.2	264	38
*H3F3A*	cattle	16.7	0.1	12.5 10⁴	0.7 10⁴
	goat	16.3	0.2	14.9 10⁴	2.3 10⁴
	sheep	16.2	0.1	19.4 10⁴	1.6 10⁴
*ACTB*	cattle	15.1	0.3	24.5 10⁴	4.3 10⁴
	goat	15.4	0.1	32.9 10⁴	2.8 10⁴
	sheep	15.5	0.2	30.2 10⁴	4.6 10⁴
*PPIA*	cattle	18.4	0.2	58.0 10⁴	0.6 10⁴
	goat	17.7	0.2	75.6 10⁴	0.8 10⁴
	sheep	17.7	0.1	65.8 10⁴	0.4 10⁴
*YWHAZ*	cattle	18.8	0.5	70.9 10⁴	1.9 10⁴
	goat	23.5	0.3	647	120
	sheep	23.6	0.2	482	81

Abbreviations for cytokine and reference genes, see Table 1.
Absolute quantification was checked by amplification of cDNA synthesized from Tempus^TM total RNA from three different animals of each species. Absolute quantification used the standard curve method and is expressed as the number of copies (nc) and Cq with standard deviation (SD) from six amplification values.

ISSN: 1746-6148