Identification and validation of reference genes for quantitative RT-PCR normalization in wheat

BMC Molecular Biology

Table 3 Coefficient of Variation^a (CV) and C_t difference (ΔC_t = C_tmax-C_tmin) determined on raw C_t values obtained by 18 primer pairs which amplified specific or multiple (m) gene transcripts of four gene families (Actin, α-tubulin, Translation elongation factor and ADP-ribosylation factor) in five data-sets.

Unigene cluster	Gene annotation	I Data set^b		II Data set^c		III Data set^d		IV Data set^e		V Data set^f
		CV	ΔC_t	CV	ΔC_t	CV	ΔC_t	CV	ΔC_t	CV	ΔC_t
Ta54225	Actin	3.87	3.01	4.14	2.90	1.83	1.21	1.99	1.14	2.95	1.57
Actin (m)	Actin	4.41	3.78	4.46	3.78	3.87	1.90	1.27	0.65	3.12	1.76
Ta1868	Actin	5.37	4.81	5.87	4.81	2.10	1.26	3.28	2.36	2.67	1.96
Ta4344	Actin	6.09	6.15	6.90	6.15	1.73	1.12	4.35	2.58	6.71	4.97
Ta20863	Actin	6.65	6.50	7.26	6.65	3.03	2.40	3.81	2.43	7.56	6.42
Ta53908	Actin	7.20	6.90	7.92	6.90	2.67	1.94	3.70	2.23	8.20	6.21
Ta25534	α-tubulin 5	4.65	4.14	4.79	4.14	3.16	2.25	2.75	1.90	2.98	2.31
Ta54519	α-tubulin 4	6.94	7.33	7.83	7.33	4.03	2.82	5.65	3.70	7.04	5.38
α-tubulin (m)	α-tubulin	7.01	6.42	7.82	6.42	3.31	1.63	3.84	1.61	8.25	5.41
Ta53981	α-tubulin 2	7.54	6.39	8.31	6.39	4.00	2.04	3.52	1.93	9.21	6.25
Ta33558	α-tubulin 3	7.77	7.02	8.59	7.02	4.09	2.56	5.21	3.52	9.37	7.02
Ta5566	α-tubulin 1	8.05	8.49	8.82	8.49	4.84	2.73	5.42	3.58	8.04	6.60
Ta53964	Translation EF 1α-subunit	3.55	3.46	3.74	3.46	1.04	0.66	1.74	0.91	2.71	2.00
TEF-1α (m)	Translation EF 1α-subunit	4.53	4.09	5.08	4.09	2.20	1.17	2.77	1.49	4.01	2.60
Ta659	Translation EF 1α-subunit	4.82	4.35	5.27	4.35	2.12	1.16	2.50	1.22	3.82	2.48
Ta2291	ADP-ribosylation factor	4.54	3.46	4.85	3.46	2.02	1.27	1.38	0.70	3.86	2.55
Ta45379	ADP-ribosylation factor	4.59	4.03	4.85	4.03	2.21	1.35	2.15	1.18	5.16	3.77
ADP-RF (m)	ADP-ribosylation factor	6.70	4.68	6.24	4.68	3.73	2.23	2.22	1.19	6.26	4.34

^aCV = Standard deviation divided by mean and multiplied by 100 of MST (Mean Square Total) among tissues, biological experiments and technical replicates within each of the two experiments.
^bAll the 24 samples including tissues, developmental stages and temperature treatments.
^c18 tissues and developmental stages.
^dSix samples consisting of two temperature treatments (4°C and 33°C) for 24 and 48 h and their controls.
^eSix floral organs from fully emerged spikes.
^fSix vegetative tissues and developmental stages (shoots, stems and leaves).

ISSN: 1471-2199