Genotype analysis:
Genomic DNA was extracted from peripheral blood leukocytes of the
proband as well as his parents. Genotype analysis was performed on
extracted DNA using specific primer sets for all seven exons of CTSC
gene(Moghaddasian et al., 2014). Both strands of PCR products were
directly sequenced using ABI Prism 310 Automated Sequencer and analyzed
with the DNA sequence assembly software (Sequencher 4.10.1, Gene Codes
Corporation). Numbering of CTSC nucleotides and amino acids was in
accordance with the reference sequence on Gene Bank (NM_001814). Fifty
healthy volunteers were also analyzed as a control group.