Markers for distinguishing Orostachys species by SYBR Green-based real-time PCR and verification of their application in commercial O. japonica food products

Applied Biological Chemistry

Table 3 Target genes, primer sequences, amplicon sizes, and T_m values for the Orostachys species-specific qPCR developed in this study

Target species	Target gene/region	Primer	Length (bp)	Sequence (5′ → 3′)	Size (bp)	T_m (°C)
Plant system (positive control)	18S rRNA	18S rRNA_F	25	TCTGCCCTATCAACTTTCGATGGTA	137	58
Plant system (positive control)	18S rRNA	18S rRNA_R	25	AATTTGCGCGCCTGCTGCCTTCCTT	137	58
O. japonica	trnT-L	trnT-L_F	24	GAAACTACAGAAAGAAAGGATGAA	140	61
	trnT-L	trnT-L_R	18	CTCGGAATCGCTTCCTAC	140	61
	ndhF	ndhF_F	20	CATCTATAAATCTTTTACAG	149	51
	ndhF	ndhF_R	20	TTTTTGAAAGATGAATAAAC	149	51
	rpoB	rpoB_F	21	ATCGCTCACATATTCTTCTGA	116	58
	rpoB	rpoB_R	20	CGCGAGAATTCATTAGAGGT	116	58
O. iwarenge and O. malacophyllus	matK	matK_F	19	CGAATCCATACTCGGTTTT	129	58
	matK	matK_R	20	TGAATAGAAAAGCCTTCTAG	129	58
	trnS-G	trnS-G_F	20	GTGATTTTTATCCAAATTTG	155	55
	trnS-G	trnS-G_R	21	GATTTAGTTACGATTAGAAAG	155	55
	trnQ-rps16	trnQ-rps16_F	18	TCACCTAGTGCATCTCGG	102	62
	trnQ-rps16	trnQ-rps16_R	23	GGTTTCATAGAACAGATCAAGGT	102	62
	ycf2	ycf2_F	18	TGTGGGGCTAATAGTTTG	181	58
	ycf2	ycf2_R	19	TAACCAGGAACTTGTTCAG	181	58
O. latiellipticus and O. ramosus	matK	matK_F	19	AGGTACGCCTCTCCTGATA	186	62
	matK	matK_R	23	TCTAATAGTTGACTCCGTACAAA	186	62
	trnT-L	trnT-L_F	22	GCATGTTATGTTTCTCATTCAC	156	60
	trnT-L	trnT-L_R	20	ACTTGAGGCTATGTCAATTC	156	60
	trnQ-rps16	trnQ-rps16_F	19	ATTTGGGATTTAAATAGGG	115	54
	trnQ-rps16	trnQ-rps16_R	20	AGTACTCCTTCTATAGTTAG	115	54
	ITS	ITS_F	18	GAAGCGAAAATCGGACAT	152	59
	ITS	ITS_R	19	GCCATTCACACCAAGTATC	152	59
O. margaritifolius	ndhF	ndhF_F	18	AAATTCCTCTGAAACTTG	218	53
O. margaritifolius	ndhF	ndhF_R	18	TTATAGTACTTCCCCTAG	218	53

Nucleotides in underlining letters indicate mismatched sequences in the designed primers