Sanger sequencing of genomic DNA and genotyping of parent-offspring trios
Direct bidirectional Sanger sequencing of CACNA1HP1523-R1584 from genomic DNA of indicated subjects was performed following PCR amplification using primers
CACNA1H_25F (5′-GACCCACCGCCTCTGTG-3′) and CACNA1H_25R (5′-AGCGCCTTACTCCTGCG-3′).
Parent-offspring trios were genotyped as previously described, except for locus D7S820 in kindred 1390 (primers [5′-ATGTTGGTCAGGCTGACTATG-3′] and [5′-GATTCCACATTTATCCTCATTGAC-3′]) (Scholl et al., 2013). Alleles without known frequencies in the population were omitted from the analysis.