Designing universal primers for the isolation of DNA sequences encoding Proanthocyanidins biosynthetic enzymes in Crataegus aronia

Table 1 Characteristics of the designed primers specific for Rosaceae PAs biosynthesis genes used in this study

Gene name	Primer	Primer sequence (5' → 3')	T_m°C^a
ANR	ANRFwd1	CCCACCTMACAGCACTACAA ^b	53.8
	ANRRev1	TACCGACCAGAAGCAGATTC	51.8
	ANRFwd2	GGAGTGATTTGGAGTTCTTG	49.7
	ANRRev2	GGAAAATCTCCAAACTCAGT	47.7
ANS	ANSFwd1	TTTGAYCTTCCCATTGAGCA	49.7
	ANSRev1	GCATTTTGGGTAGTAGTTGA	47.7
	ANSFwd2	CAGCTTGAGTGGGAGGAYTA	53.8
	ANSRev2	GCAGGCCRGGAACCATGTTG	57.9
4-CL	4-CLFwd1	CCTGGAGAGATTTGCATCAG	51.8
	4-CLRev1	AATACTCGATTTATTCTTTTATA	42.8
	4-CLFwd2	ATCATGAAAGGTTAYCTTAATG	45.5
	4-CLRev2	GGAATGGCTTCGATGAAAAA	47.7
CHI	CHIFwd1	AGGGGGTYGGAGATTCAGGG	55.9
	CHIRev1	GGGAAGKTTTGATCYTTGAA	45.6
	CHIFwd2	TTCGTGAAGTTCACGGCGAT	51.8
	CHIRev2	TGGGAAGGTCTGATCTTTGA	49.7
C4H	C4HFwd1	CTCCGYATGGGRCAGCGCAA	60
	C4HRev1	TTGTTGTACATCATMAGCTG	47.7
	C4HFwd2	CAGCTBATGATGTACAACAA	47.7
	C4HRev2	TGGATTTCRGGGTGGTTCAC	53.8
CHS	CHSFwd1	AAGGCCATYAAGGAATGGGG	53.8
	CHSRev1	AATGTRAGCCCDACTTCACG	53.8
	CHSFwd2	AAGGAATGGGGHCAGCCCAA	55.9
	CHSRev2	CTCACYTCRTCCAAAATAAA	47.7
DFR	DFRFwd1	CCTGACGCTGTGGAAGGCGG	60
	DFRRev1	TAATGGTRATGAAATCAATG	43.6
	DFRFwd2	TGCASCGGAGTGTTYCATGT	53.8
	DFRRev2	TGAGGCTTGGTGGCATRGATGG	58.6
LAR	LARFwd1	GTTCGTMGCYGAAGCCAGCC	60
	LARRev1	TGGAACYGATCCAACGGTGG	55.9
	LARFwd2	AGGGCYRATCCGGTKGAACC	60
	LARRev2	AGGGTSCGRCCAATTTTCTT	51.8
F3H	F3HFwd1	GACATGTCCGGYGGCAAAAAGGG	60.6
	F3HRev1	CTCATCTTCTTCTTGTACAT	45.6
	F3HFwd2	ACGTGGATCACCGTTCAACC	53.8
	F3HRev2	GGTTGAACGGTGATCCACGT	53.8
PAL	PALFwd1	ATCGATGTTTCRAGGAACAA	47.7
	PALRev1	AAAGAGTTAACATCTTGGTT	43.6
	PALRev2	AAAAATGTRGAAGACATGAG	45.6

^a Melting temperature.
^b Degenerate nucleotides codes where B (C/G/T), D (A/G/T), H (A/C/T), K (G/T), M (A/C), N (A/C/G/T), R (A/G), S (G/C), V (A/C/G), W (A/T), Y (C/T).

ISSN: 1756-0500