How to perform RT-qPCR accurately in plant species? A case study on flower colour gene expression in an azalea (Rhododendron simsii hybrids) mapping population

Table 1 Target genes

Code	Gene	Acc. No.	Primer (5′-3′)	Ampl.	Position
ANS	anthocyanidin synthase	AB289596	CCAAGAATCCGTCCGACTACA	65 bp	Exon1/2
ANS	anthocyanidin synthase	AB289596	GGTTAGGCCTCTCAGGTGCTT	65 bp	Exon1/2
CHS	chalcone synthase	AJ413277	TGGGAATCAACGGTTTTGGAA	151 bp	Exon1
CHS	chalcone synthase	AJ413277	CTCGGGCTTAAGGCTCAACTT	151 bp	Exon1
DFR	dihydroflavonol 4-reductase	AJ413278	CGTCATGAGGCTGCTTGAAC	151 bp	Exon1/2
DFR	dihydroflavonol 4-reductase	AJ413278	AAAGCTCCCTTCCTCGTTGAG	151 bp	Exon1/2
F3H	flavanone 3-hydroxylase	AB289594	GGGCTCCAGGCCACTAGAG	87 bp	Exon2
F3H	flavanone 3-hydroxylase	AB289594	ATGGTCGCCCAAATTGACAA	87 bp	Exon2
F3′H	flavonoid 3′-hydroxylase	AB289597	AAGAGCTGGACTCAATTGTTGGA	87 bp	Exon3
F3′H	flavonoid 3′-hydroxylase	AB289597	CCTTGATGATGGCTTGGAGGTA	87 bp	Exon3
FLS	flavonol synthase	AB289599	CAAGGATGTCATGGGCTGTGT	75 bp	Exon3
FLS	flavonol synthase	AB289599	CGTTAATGAGCTCCGGAATAGG	75 bp	Exon3

Primer pairs for target genes were developed using Primer Express 2.0 (Applied Biosystems). EMBL accession numbers and the length of the amplicons are indicated. The position of the amplicons at the genomic DNA level is marked.

ISSN: 1471-2199