Table 3.

SNP locations in the mitochondrial COI gene and SNaPshot extension primer sequences used in this study. The size in terms of nucleotide bases and the optimal primer concentration in the SNaPshot multiplex are also shown.

SBE system	Location	SBE primer sequence (5′ → 3′)	Primer size (nucleotides)	[Opt.‡] (μM)
Calliphoridae	CA90	CTTGATCNGGAATARTHGGAACTTC∗	25	0.80
		CTTGATCCGGTATAATYGGAACTTC∗†	25	0.01
		CTTGATCAGGAATAATTGGTACTTC†	25	0.20
	CA72	TTTTTTACTTTATAYTTTATTTTTGGAGCTTGATC∗	35	0.04
		TTTTTTACTTTATATTTYATTTTCGGAGCTTGATC∗†	35	0.06
	CA168	TTTTTTTTTTTTTTTTGGAGAYGAYCAAATTTATAATGTAATTGT∗	45	0.10
	CA261	TTTTTTTTTTTTTTTTTTTTTTTTTTAATTGATTAGTTCCWTTAATACTAGGRGC∗	55	0.20
		TTTTTTTTTTTTTTTTTTTTTTTTTTAATTGAYTAGTTCCTTTAATGTTAGGAGC∗†	55	0.10
		TTTTTTTTTTTTTTTTTTTTTTTTTTAATTGATTAGTYCCTTTAATATTAGGAGC∗†	55	0.13
	CA252	TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTGGAGGRTTTGGAAATTGAYTAGTYCCWTTAAT∗	65	0.14
	CA243	TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTGGAGGRTTYGGWAATTGAYTAGT∗	75	1.80
Sarcophagidae	SA1491	AATTCAGAATAACTATGTTCAGCTGG	26	0.06
	SA1488	TTTTTTTTTTTTTGAATAACTATGTTCAGCTGGHGG∗	36	0.15
	SA1479	TTTTTTTTTTTTTTTTTTAACTATGTTCAGCTGGHGGDGTRTTTTG∗	46	0.30
	SA1485	TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTCTATGTTCAGCTGGHGGDGT∗	56	0.40

^∗Degenerated primers were used to detect target SNPs based on IUPAC nucleic acid sequences; ^†additional primers for samples with low peak signal sites; ^‡[Opt.]: optimal concentration of primer.