Development and characterization of microsatellite markers for diploid populations of the wind-pollinated herb Mercurialis annua

Table 1 Characteristics of the three primer multiplexes developed for M. annua, tested on 100 individuals

M	Locus	Primer sequences (5′–3′)		Repeat motif	Size range (bp)	Ta (°C)	FC (µM)	N_A	H _O	H _E	Error rate %
1	1101	F	5′FAM—ACTAAAGTCCACTCATGGATGC	(TCTT)₉	94–115	60	0.1	6	0.580	0.599	1.8
	1101	R	GTATCCGTCCCTGGAGGAAG	(TCTT)₉	94–115	60	0.1	6	0.580	0.599	1.8
	661	F	JOE—CAACGCTTTTGTACAGGGGG	(CTAT)₇	187–203	60	0.1	6	0.620	0.685	0.0
	661	R	ACATTTATCCCTCCCCAGGC	(CTAT)₇	187–203	60	0.1	6	0.620	0.685	0.0
	7575	F	AT550—AGTCACAGCTTGCAAATCTCC	(TACA)₇	222–255	60	0.1	12	0.690	0.851	6.8
	7575	R	TGCATCTTCTGGTTTCATCCC	(TACA)₇	222–255	60	0.1	12	0.690	0.851	6.8
2	317	F	5′FAM—TTTACCGAGATGCTCCCCAG	(CTAT)₇	188–273	60	0.2	11	0.760	0.804	1.9
	317	R	AAAGCCCCTGGAATTGTTGC	(CTAT)₇	188–273	60	0.2	11	0.760	0.804	1.9
	13654	F	JOE—TGCTGTTGGGATAAACAGGC	(TCTT)₉	191–219	60	0.2	7	0.730	0.792	1.1
	13654	R	CACTGCAAACGCATGAATCG	(TCTT)₉	191–219	60	0.2	7	0.730	0.792	1.1
	4048	F	AT550—AGGGGTACCGTTTTGAGTCC	(GT)₁₂	212–240	60	0.2	8	0.570	0.778	5.9
	4048	R	CGATCCCTGCACAATCTGAC	(GT)₁₂	212–240	60	0.2	8	0.570	0.778	5.9
3	330	F	HEX—TAGCACGGAAACAAAAACGC	(AAAG)₈	168–196	59	0.2	9	0.670	0.769	0.8
	330	R	CTCGGTGGATTACCAAGACG	(AAAG)₈	168–196	59	0.2	9	0.670	0.769	0.8
	3889	F	5′FAM—TCGGTTTGATTTTTGAACACCC	(TCTA)₇	216–242	59	0.2	7	0.710	0.696	0.0
	3889	R	CACTGTAATCATGCCAAAGCTG	(TCTA)₇	216–242	59	0.2	7	0.710	0.696	0.0

Fluorescent tags attached to the 5′ end of the forward primers and are indicated in italic
M, multiplex; Ta, annealing temperature; FC, final concentration of each set of primers in the PCR reaction; N_A, number of alleles; H _O, observed heterozygosity; H _E, expected heterozygosity

ISSN: 1756-0500