Table 1.

Primers used for amplifying and Sanger sequencing exonic and flanking regions of canine NAGLU gene and for genotyping the causative insertion and wild-type alleles in Schipperkes.

Exon #	Exon length (bp)	Forward Primer	Reverse Primer	Amplicon Length (bp)	Annealing Temperature (°C)
1	424	ATGTGAAAGCTCTCCAGGTACA	CGATGTCACCGTTTCCATTCTTC	925	66
2	148	GTGAGTCCTGGAGTGAAACAGT	TAGCGTTTCTAGTGAGGTGCTG	485	65
3–4	147 (Exon 3) 86 (Exon 4)	TTGCAACAAAGCTGACCCATTAG	GCTGCCATTTGCTAAGACTGTG	798	67
5	257	CACTGCTCCATCTAGGACTCTG	AGTGCTTGGTCAACTGTCAAGG	682	67
6a	1450	GACAACACTGCCCTAGAGATCC	CCTCGCCTCCACATAGTACAAG	953	67
6b		ATGGTTACCACTGTCTGGTACA	AAACGTATTGGGAGAGGATTCCC	1043	65
Genotyping		GCATTCCCTTCCAACAGCACCAGT	GCCCACAAGGAGCCAGCCACCAAT	169	68

Primer pairs 6a and 6b were designed to amplify and sequence exon 6 to make it across this relatively long exon.