Development of a cost-effective high-throughput process of microsatellite analysis involving miniaturized multiplexed PCR amplification and automated allele identification

Table 3 Microsatellite markers size ranges and primer design and concentrations in multiplex PCR reactions

PCR reaction	Marker	Range of PCR product size (bases)	Dye	Primer sequence (5^′→3^′)	Primer (μM)
1	BAT25	130	Fam	FamTCGCCTCCAAGAATGTAAGTgtgtcttTCTGCATTTTAACTATGGCTC	0.25
1	BAT26	120	Hex	HexTGACTACTTTTGACTTCAGCCgtgtcttAACCATTCAACATTTTTAACCC	2.00
1	D3S3623	207–223	Fam	FamCCATGTTGGTTAAAGGCAAGgtgtcttCTCTGAACTGAAGTGACCTCC	0.25
1	D6S262	167–183	Hex	HexATTCTTACTGCTGGAAAACCATgtgtcttGGAGCATAGTTACCCTTAAAATC	0.50
1	D7S481	181–199	Ned	NedTTCTCATTCTCACCCCCATgtgtcttATCCCCCACTGTCTCCAA	0.50
2	D2S123	203–233	Hex	gtgtcttAACAGGATGCCTGCCTTTAHexGGACTTTCCACCTATGGGAC	1.00
2	D3S1262	132–146	Hex	HexCAGTTGTGAGCCACCATGTCgtgtcttCAGTTTTTATGGACGGGGT	2.00
2	D9S171	102–122	Ned	NedGTGAACCTCATCTCTGTCTGCTgtgtcttACTTTATTAACAATCAGTATTTTCC	0.25
2	D17S250	188–203	Ned	gtgtcttGTAAGCATAAAAAGGAAGAATCANedTTACAGGCATGAGCCACTC	0.50
2	D18S61	150–176	Fam	FamATTTCTAAGAGGACTCCCAAACgtgtcttGAAACTCAGGAGCATGGTTATG	0.50

Summary of microsatellite markers, population size ranges for amplified products with specific primer sets, and primer sequences, labels, and concentrations used in multiplex PCR amplifications.

ISSN: 1479-7364