Simple allele-discriminating PCR for cost-effective and rapid genotyping and mapping

Table 2 Primer sequences for three different alleles

SNP	Primer	Direction	Sequence (5'-3')
lug-16	WT-Specific	Reverse	CCACCAGGTGCGTCAATATC
	Mutant-Specific	Reverse	CCACCAGGTGCGTCAATATT
	Common	Forward	TTGTATGCAAGTATGTGACTTTA
*lug-16 (Amplifluor)**	WT-FAM	Reverse	GAAGGTGACCAAGTTCATGCT TCCACCAGGTGCGTCAATATC
	Mutant-JOE	Reverse	GAAGGTCGGAGTCAACGGATTT CCACCAGGTGCGTCAATATT
	Common HT	Forward	CTGCAGTTGCTCTGTTTCCTAA
luh-1	WT-Specific	Forward	GGAGGGTTTCTTTTTGAGTTG
	Mutant-Specific	Forward	TGGAGGGTTTCTTTTTGAGTTA
	Common	Reverse	CCATGATGGTTTGTTGCTGAT
lug-3	WT-Specific	Reverse	TTGATGTTGTTGTTGCTGCGG
	Mutant-Specific	Reverse	TTGATGTTGTTGTTGCTGCCA
	Common	Forward	ACTAAGCTGGAGTATTTCTATTT

*: The underlined sequences are 5' extended primer sequences that specifically pair with the 3' region of the FAM or JOE universal fluorescent primers, respectively of the Amplifluor SNPs Genotyping System.

ISSN: 1746-4811