Table 3. Ct values of original assays and homogenous assays targeting on the SNPs in five wheat lines with SNPs.

Real-time PCR assay	Primers and probe	Wheat cultivar	Ct Values
			50 ng	5 ng	1 ng
ALMT1	Q-ALMT1-F/R and Q-ALMT1-p	Zhongguochun (A>C)	29.49	32.95	35.25
		Zhengmai 9023 (G>A)	29.34	32.89	35.93
		Cultivars without SNPs	27.36-29.21	30.77-32.65	32.72-35.74
	Q-ALMT1-F2/R and Q-ALMT1-p	Zhongguochun (A>C)	27.90	31.16	33.22
	Q-ALMT1-F/R and Q-ALMT1-p2	Zhengmai 9023 (G>A)	28.14	31.40	32.52
PKABA1	Q-PKABA1-F/R and Q-PKABA1-p	Zhoumai 16 (A>G)	30.29	33.39	35.40
		Zimai 12 (A>G)	30.27	33.54	35.41
		Cultivars without SNPs	27.66-29.93	30.66-32.66	32.49-34.86
	Q-PKABA1-F/R and Q-PKABA1-p2	Zhoumai 16 (A>G)	28.60	31.70	34.17
		Zimai 12 (A>G)	28.91	32.17	34.64
Acc1	Q-Acc1-F/R and Q-Acc1-p	Cannizzo wheat (C>T)	26.29	29.48	31.78
		Cultivars without SNPs	22.77-24.67	25.97-28.49	27.83-30.5
	Q-Acc1-F2/R and Q-Acc1-p	Cannizzo wheat (C>T)	24.39	27.78	29.19