Reference genes identification for normalization of qPCR under multiple stresses in Hordeum brevisubulatum

Table 1 Comprehensive details of 11 candidate reference genes used for normalization

Gene symbol	Primers (5′–3′) Forward/reverse	Length (bp)	T_m (°C)	PCR efficiency	Regression coefficient (R²)
ACT	TGCATGGTGTTCTCTGCACT GCCAAAGCGTGATACTGTCG	121	60	0.92	0.997
ADP	CAAGAGATAGTATGTGTGCGTATG AACCACGGCACAGAAATACTGAT	115	60	1.03	0.997
CYP2	CCTGTCGTGTCGTCGGTCTAAA ACGCAGATCCAGCAGCCTAAAG	151	60	0.96	0.991
EF-1α	CAACAACGCCCAGGAACAAC AGAAGAAGGACCCCACTGGT	159	60	0.94	0.994
GAPDH	AGCTGCACCACTAACTGCCT AACAGTGGTCATCAAACCCTCAAT	84	60	0.96	0.997
HSP90	CAGCACCTCCTTGATGACCTT GGAGTTTGAGGGCAAGAAGC	136	60	0.97	0.997
TUBα	CGAGATGCACTCCCTCATGG CGTCTTCGTACTCGCCTCTC	128	60	1.15	1.000
TUBβ6	CCCGACGAGAACGCTTCAAT CTGGATGTGCAGGATCTCCC	74	60	0.89	0.996
UBI	TGGATGTTGTAGTCGGCGAG ACGTCAAGGCCAAGATCCAG	112	60	0.90	0.996
18SrRNA-1	CCCGCTTCTAAGTCGGTGTT CAACAGAAGCGCGATACAGC	99	60	0.88	0.997
18SrRNA-3	TTTCGTGAGGGCCTGCTTAG GACTCACAGAACATGGGGCA	83	60	0.97	0.997

The correlation coefficients (R²) and slope values were obtained from the standard regression curves and the PCR amplification efficiencies (E) were calculated according to the following equation: E = (10^−1/slope − 1)

ISSN: 1746-4811