A duplex, SYBR Green I-based RT-qPCR assay for the simultaneous detection of Apple chlorotic leaf spot virus and Cherry green ring mottle virus in peach

Table 1 Primer sequences and amplicon characteristics for PCRs

PCR type	Name	Locus description	Primer Sequence (5’-3’)	Product size (bp)	Product TM (°C)	RT-qPCR Efficiency (%)	R²	Concentration (nM)
qPCR^a	AC62F	Coat protein	AAATACCCGGAGCTGATGTTTG	138	79.40 ± 0.24	97.0	0.9989	250
	AC62R	Coat protein	CTTCGCCTCATTTTCACTCTTTG	138	79.40 ± 0.24	97.0	0.9989	250
	CG732F	Coat protein	CAATTCAGGAGACGAACCCAG	181	81.44 ± 0.37	101.0	0.9939	250
	CG732R	Coat protein	TTCCCGACCATCTTTGTTTTG	181	81.44 ± 0.37	101.0	0.9939	250
	RPIIF	RNA polymerase subunit	TGAAGCATACACCTATGATGATGAAG	128	79.41 ± 0.29	93.6	0.9996	250
	RPIIR	RNA polymerase subunit	CTTTGACAGCACCAGTAGATTCC	128	79.41 ± 0.29	93.6	0.9996	250
	UBQ10F	Ubiquitin 10	AAGGCTAAGATCCAAGACAAAGAG	146	84.36 ± 0.12	98.5	0.9958	500
	UBQ10R	Ubiquitin 10	CCACGAAGACGAAGCACTAAG	146	84.36 ± 0.12	98.5	0.9958	500
nPCR^b	AC84F	Coat protein^c	TCTGCAAGAGAATTTCAGTT	777	\	\	\	\
	AC84R	Coat protein^c	GTCTACAGGCTATTTATTATAAG	777	\	\	\	\
	CG94F	Coat protein^d	CCTCATTCACATAGCTTAGGTTT	929	\	\	\	\
	CG94R		ACTTTAGCTTCGCCCCGTG		\	\	\	\

^aquantitative PCR; ^bnormal PCR; ^cgenomic location 6735–7512, GenBank accession JN634761; ^dgenomic location 7306–8235, GenBank accession JX501671.

ISSN: 1743-422X