Quantitative real-time PCR with SYBR Green detection to assess gene duplication in insects: study of gene dosage in Drosophila melanogaster (Diptera) and in Ostrinia nubilalis (Lepidoptera)

Table 1 Primers used for the qPCR amplifications.

Insect	Target gene	Primer pair	Primer name	Sequence (5'→3')
D. melanogaster	RpS3	s3-1	s3-1 F	TCTTTCTTTTCTGCGCACCA
			s3-1 R	TCGCATTCATTTTGACGTCG
	BarH1	h1-4	h1-4 F	CCAGGACGATCCGTTGACA
			h1-4 R	GATCTGATCCTCGTCGTCCG
		h1-5	h1-5 F	TCCAGGTGTGAGCGGTACG
			h1-5 R	ATTAGCACACGCACACAATCG
	BarH2	h2-4	h2-4 F	CAGAACCAAATGGAAGCGTCA
			h2-4 R	TCGGCCAGCAGTTCCAAG
		h2-7	h2-7 F	GGAGGAACTGGCCCTGGA
			h2-7 R	GGAGGGTTGAATTCTCTCGGA
O. nubilalis	RpS3	s3-1	s3-1 F	GAGCTACTGGGAGAGAAGG
			s3-1 R	GATGTTGAAACGCTTCTGGA
		s3-2	s3-2 F	GCGTTTCAACATCCCTGAACA
			s3-2 R	GGCGACTTTCTCAGCGTACAG
	tpi	t1	t1 F	GGCGACAAGAATCAAATCAATG
			t1 R	AGGACCCTTTTTCAGAGTGTTCAC
	ldh	l1	l1 F	GAATAAATCGGGCTCGAAGGAC
			l1 R	TCACGCGAGACAGCTTCAAA
	cadherin	e26	e26F	ACGGCAACAACGAGGGTCT
			e26R	GAGATGACGTTGCGCGACT
		e28	e28F	CGAGCCACACAGAAGACGAC
			e28R	TGCTCGCACGGTCTATGATG

The letters F or R at the end of the primer names refer to their respective orientation (F = forward; R = reverse). The qPCR product sizes were 51 base pairs in all reactions except for primer pairs s3-1 and e26 of O. nubilalis, with lengths of 70 and 52 base pairs respectively.

ISSN: 1756-0500