PCR-based genotyping assays to detect germline APC variant associated with hereditary gastrointestinal polyposis in Jack Russell terriers

Table 1 Primers and probes used for PCR-based assays for detection of the germline APC variant

			Sequence		Product size (bp)
Primers used for PCR-direct sequencing
Primers	sense	5′-	AGTCCCACCTTCAAAAATCC	−3’	385
Primers	antisense	5′-	AACTAAAAATGCAATTATCTTGAATG	−3’	385
Primers used for PCR-RFLP and PCR-directsequencing of FFPE samples
Primers	sense	5′-	TCTTTTGGCATTGTGTAAACTTG	−3’	156
Primers	antisense	5′-	CTTACATTTTCAGTTAAAGGGAGACT	−3’	156
Primers and probes used for Taq-Man duplex real-time PCR assay
Primers	sense	5′-	TTTAGTGAGATTCTGAAGTTGAGCATAATA	−3’	78
Primers	antisense	5′-	TCATTACTTCTTGCTGATCTTGACAA	−3’	78
Probe for Wild-type allele		5′-	(VIC)- CAATCTTTTTCCTTTTC-(MGB)	−3’
Probe for Mutant allele		5′-	(FAM)-CAATCTTAATCCTTTTC-(MGB)	−3’

VIC 2-chloro-7’-phenyl-1,4-dichloro-6-carboxy-fluorescein, FAM 6-carboxyfluorescein, NFQ nonfluorescent quencher, MGB minor groove binder

ISSN: 1746-6148