Validation of reference genes for gene expression analysis in olive (Olea europaea) mesocarp tissue by quantitative real-time RT-PCR

Table 3 BestKeeper descriptive statistical analyses of eight reference genes in Barnea, Frantoio and Picual mesocarp together (A) and individually (B, C and D)

Factors	Stability of reference genes in Barnea, Frantoio and Picual (A)
	60S RBP L18-3	PP2A	PTB	GAPDH	EF1-alpha	OUB2	TIP2	TUBA
n	24	24	24	24	24	24	24	24
SD [±Cq]	0.9	1	0.93	0.94	0.99	1.07	2.64	1.54
CV [% Cq]	2.87	3.08	2.69	3.29	4.26	4.41	7.66	5.11
r value	0.654	0.805	0.683	0.717	0.769	0.711	0.832	0.665
p value	0.001	0.001	0.001	0.001	0.001	0.001	0.001	0.001
Ranking	5	1	4	3	2	7	6	8
	Stability of reference genes in Barnea (B)
n	8	8	8	8	8	8	8	8
SD [±Cq]	1.51	0.94	1.56	0.99	0.98	1.00	3.50	1.50
CV [% Cq]	4.86	3.49	4.56	4.31	4.29	4.45	10.12	4.91
r value	0.744	0.886	0.720	0.796	0.854	0.771	0.866	0.811
p value	0.034	0.003	0.044	0.039	0.007	0.023	0.005	0.014
Ranking	5	1	6	3	2	4	7	8
	Stability of reference genes in Frantoio (C)
n	8	8	8	8	8	8	8	8
SD [±Cq]	0.53	0.47	0.26	0.68	0.44	0.28	1.15	1.38
CV [% Cq]	1.68	1.46	0.75	2.36	1.95	1.19	3.32	4.56
r value	0.778	0.686	0.713	0.835	0.827	0.796	0.510	0.682
p value	0.004	0.021	0.032	0.010	0.004	0.020	0.003	0.002
Ranking	4	6	5	1	2	3	8	7
	Stability of reference genes in Picual (D)
n	8	8	8	8	8	8	8	8
SD [±Cq]	0.36	0.53	0.83	0.98	0.99	0.96	3.32	1.51
CV [% Cq]	1.13	1.64	2.38	2.56	2.80	2.42	9.66	5.04
r value	0.969	0.883	0.659	0.797	0.809	0.946	0.867	0.753
p value	0.001	0.004	0.015	0.018	0.015	0.001	0.005	0.031
Ranking	1	3	6	5	4	2	7	8

Abbreviations: n number of samples, SD [±Cq] standard deviation of Cq values, CV [% Cq] coefficient of variance expressed as percentage of Cq values, r coefficient of correlation, p probability value. Genes showing the highest r value and genes with SD > 1 have been highlighted in bold. Ranking of the eight reference genes based on their r-value and SD values have also been shown.

ISSN: 1756-0500