In terms of genotype analysis, GLA gene sequence analysis was performed. The seven exons of the GLA gene were amplified by polymerase chain reaction (PCR) with specific primers and sequenced by the Sanger method on a genetic analyzer (Applied Biosystems Inc. CA, USA). Results were analyzed using the software SeqScape 2.5.0 (Applied Biosystems Inc. CA, USA). DNA was extracted with an QIAamp DNA Blood Mini Kit (Qiagen Inc.). A total of 7 pairs of PCR primers were designed to amplify the 7 exons encoding the GLA gene (29 (link)). The PCR amplifications were carried out using Taq DNA polymerase (PhireII HS, Thermo Inc.) and a PCR protocol was set, having an initial hold of 1 minute at 95°C, 45 cycles (of 10 seconds at 95°C, 10 seconds at 60°C and 20 seconds at 72°C), and a final extension of 1 minute at 72°C. After the thermal cycle protocol for PCR, the product was checked using 2% agarose gel electrophoresis. PCR products were purified using the ZR-96 DNA Sequencing Clean-up Kit (Zymo Research Corp.) and the purified products were sequenced bidirectionally on a ABI 3130 capillary gel electrophoresis system (Applied Biosystems Inc. CA, USA) according to the manufacturer’s protocol. The exons of the gene and the exon-intron connections were analyzed by the SeqScape 2.5.0 (Applied Biosystems Inc. CA, USA) software and the sequence variations were determined.