Betaine

To determine the genotype frequency of PICK1-rs713729, PICK1-rs2076369 and BDNF-rs6265 an ARMS-PCR method was used. Speci c primers for PCR ampli cation were designed via web tools, such as Primer1 and also WASP (web-based allele-speci c primer designing tool) [28] .
PCR ampli cations for PICK1-rs713729, PICK1-rs2076369 and BDNF-rs6265 were conducted in a 10-15 microliter (μl) volume per reaction, containing 3 µl Taq 2x master mix (Ampliqon, Germany), 10 µM of each primer and 100 nanogram (ng) DNA. Moreover, the speci c primers used to detect PICK1-rs713729, PICK1-rs2076369 and BDNF-rs6265 SNPs are listed in Table 4. For PICK1-rs2076369, we also used betaine (Ampliqon, Germany) as an enhancer in PCR.
The ARMS-PCRs condition for each primer is as follows, Table 5. In general, initial denaturation at temperature 94 °C for ve minutes, then 35 cycles including denaturation at 94 °C for 25 seconds, annealing at alternative °C for 25 seconds (based on each primer), an elongation at 72 °C for 30 seconds followed by 72 °C for seven minutes as the nal elongation step (Table 5).
Absence or presence of mutant or normal PCR products were detected via gel electrophoresis in 3% agarose gel by ultraviolet (UV) trans illuminator (Gel Doc; U:Genius). Table 4 Primer sequences used for genotyping in ARMS-PCR.
Table 5 The ARMS-PCRs condition for targeted SNPs.

To determine the genotype frequency of PICK1-rs713729, PICK1-rs2076369 and BDNF-rs6265 an ARMS-PCR method was used. Speci c primers for PCR ampli cation were designed via web tools, such as Primer1 and also WASP (web-based allele-speci c primer designing tool) [28] .
PCR ampli cations for PICK1-rs713729, PICK1-rs2076369 and BDNF-rs6265 were conducted in a 10-15 microliter (μl) volume per reaction, containing 3 µl Taq 2x master mix (Ampliqon, Germany), 10 µM of each primer and 100 nanogram (ng) DNA. Moreover, the speci c primers used to detect PICK1-rs713729, PICK1-rs2076369 and BDNF-rs6265 SNPs are listed in Table 4. For PICK1-rs2076369, we also used betaine (Ampliqon, Germany) as an enhancer in PCR.
The ARMS-PCRs condition for each primer is as follows, Table 5. In general, initial denaturation at temperature 94 °C for ve minutes, then 35 cycles including denaturation at 94 °C for 25 seconds, annealing at alternative °C for 25 seconds (based on each primer), an elongation at 72 °C for 30 seconds followed by 72 °C for seven minutes as the nal elongation step (Table 5).

To determine the genotype frequency of PICK1-rs713729, PICK1-rs2076369 and BDNF-rs6265 an ARMS-PCR method was used. Speci c primers for PCR ampli cation were designed via web tools, such as Primer1 and also WASP (web-based allele-speci c primer designing tool) [28] .
PCR ampli cations for PICK1-rs713729, PICK1-rs2076369 and BDNF-rs6265 were conducted in a 10-15 microliter (μl) volume per reaction, containing 3 µl Taq 2x master mix (Ampliqon, Germany), 10 µM of each primer and 100 nanogram (ng) DNA. Moreover, the speci c primers used to detect PICK1-rs713729, PICK1-rs2076369 and BDNF-rs6265 SNPs are listed in Table4. For PICK1-rs2076369, we also used betaine (Ampliqon, Germany) as an enhancer in PCR.
The ARMS-PCRs condition for each primer is as follows, Table 5. In general, initial denaturation at temperature 94 °C for ve minutes, then 35 cycles including denaturation at 94 °C for 25 seconds, annealing at alternative °C for 25 seconds (based on each primer), an elongation at 72 °C for 30 seconds followed by 72 °C for seven minutes as the nal elongation step (Table 5).
Absence or presence of mutant or normal PCR products were detected via gel electrophoresis in 3% agarose gel by ultraviolet (UV) trans illuminator (Gel Doc; U:Genius).