Genomic micro ax swab gravity

Genomic DNA was extracted from the buccal cells by a Genomic Micro AX SWAB Gravity (A&A Biotechnology, Poland) according to the producer’s protocol. An allelic discrimination assay on a C1000 Touch Thermal Cycler (Bio-Rad, Feldkirchen, Germany) instrument with TaqMan^® probes was applied. To discriminate the SCN9A alleles, TaqMan^® Pre-Designed SNP Genotyping Assays (Applied Biosystems, Waltham, MA, USA) (assay ID: C__29330435_10), consisting of fluorescently labelled (FAM and VIC) minor groove binder (MGB) probes and two specific primers, were used. All samples were genotyped in duplicate.

Genomic DNA was extracted from the buccal cells by a Genomic Micro AX SWAB Gravity (A&A Biotechnology, Poland) according to the producer’s protocol. All samples were genotyped using allelic discrimination assays with TaqMan^® probes (Applied Biosystems, Carlsbad, CA, USA) on a 7500 Fast Real-Time PCR Detection System (Applied Biosystems). In the COMT1 gene, four SNPs were genotyped: C/T rs4633, A/G rs4680, C/G rs4818 and A/G rs6269, while in the OPRM1 gene, one SNP was genotyped: A/G rs1799971. To discriminate the OPRM1 rs1799971 alleles, TaqMan^® Pre-Designed SNP Genotyping Assays (Applied Biosystems, USA) (assay ID: C___8950074_1_), for COMT1 rs4633, rs4680, rs4818 and rs6269 (assays ID: C___2538747_20, C__25746809_50, C___2538750_10 and C___2538746_1_, respectively) consisting of fluorescently labelled (FAM and VIC) minor groove binder (MGB) probes and two specific primers was used [16 (link),17 (link)]. All samples were genotyped in duplicate.