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

De Keyser, Ellen; Desmet, Laurence; Van Bockstaele, Erik; De Riek, Jan

doi:10.1186/1471-2199-14-13

Table 2 Reference genes

From: 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

Gene	Acc. No.	Functional annotation	Primer (5′-3′)	Ampl.
GAPDH	FN552706	Glyceraldehyde 3-phosphate dehydrogenase	TCGGAATCAACGGTTTTGGAA	151 bp
GAPDH	FN552706	Glyceraldehyde 3-phosphate dehydrogenase	CACTTGACCGTGAACACTGT	151 bp
HK5	AM932886	Histone H3	GAAACTCCCATTCCAGAGGCT	153 bp
HK5	AM932886	Histone H3	GCATGGATGGCACAGAGGTT	153 bp
HK47	AM932894	Nucleosome assembly protein	GGTATAGGATTGACAATCCCAAGG	151 bp
HK47	AM932894	Nucleosome assembly protein	CATTCAATCTCCGTCCCTATCG	151 bp
HK65	AM932901	Protein kinase regulatory subunit γ	CGGCAGTTAGGAGCTACCTCG	151 bp
HK65	AM932901	Protein kinase regulatory subunit γ	CCCTCACCGTCCACAACATAG	151 bp
HK92	FN552699	Heterotrimeric G-protein,α subunit	ATCACAGTCATCCATGCCAATG	151 bp
HK92	FN552699	Heterotrimeric G-protein,α subunit	CGCCGCCAATTTCTGATAGT	151 bp
HK96	AM932905	Expansin	AGGTTCACAATCAATGGCCAC	151 bp
HK96	AM932905	Expansin	TGTTGCTCTGCCAATTCTGC	151 bp
HK112	FN552700	3-deoxy-D-arabino-heptulo-sonate 7-phosphate synthase	CTCCTCCCTTCCTCCCAATC	152 bp
HK112	FN552700	3-deoxy-D-arabino-heptulo-sonate 7-phosphate synthase	GTAACCGTTGTGCTCCCTACAGTC	152 bp
HK129	AM932909	Protein phosphatase	TGCAAAGATCGAATGCACGA	165 bp
HK129	AM932909	Protein phosphatase	CCTGCAAACGGAACTCGAGA	165 bp
HK134	FN552701	Chlorophyll a/b binding protein CP24 precursor	CGGTTGCTCCCAAAAAGTCTT	158 bp
HK134	FN552701	Chlorophyll a/b binding protein CP24 precursor	CTCCGCTTCTCGGTACCACT	158 bp
HK156	FN552702	Cytochrome P450 mRNA	AGCCATGACCATCTTCGCTT	156 bp
HK156	FN552702	Cytochrome P450 mRNA	GGCGATGATGCAAACGAGTT	156 bp
HK164	FN552703	Chlorophyll a/b binding protein	AAAACCTCTTCTCTTGCAAACCAT	151 bp
HK164	FN552703	Chlorophyll a/b binding protein	CTTGCCGACAGACTTCCTCAT	151 bp
HK173	FN552704	Pyruvate dehydrogenase	GGTGCGAGATTGGTATTTGGA	151 bp
HK173	FN552704	Pyruvate dehydrogenase	TTGAACTCCCAAAGCCATTGT	151 bp
HK190	FN552705	Protein disulphide isomerase	CGTATCGATCATCGGCTCGT	152 bp
HK190	FN552705	Protein disulphide isomerase	CACACCACGGAGCGTAGAACT	152 bp

Primer pairs for candidate reference genes were developed using Primer Express 2.0 (Applied Biosystems) based on EST-fragments (described by their EMBL accession number). The length of the amplicons and the putative function annotated to the sequences is indicated.

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ISSN: 1471-2199

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