Efficiency of two available kits for amplification of three EGFR SNPs in patients with NSCLC: 181946 G/A (rs2293347), -191 C/A (rs712830) and -216G/T (rs712829) with GC-rich regions

Aim . To conduct a comparative analysis of two available kits that contain all necessary reagents and additives in a single reaction mixture, including 100 mM Tris-НCl, 100 mM KCL, 4 mM MgSO4, 0.2% of Tween 20, for the amplification of the three most common EGFR (epidermal growth factor receptor) gene polymorphisms in patients with non-small cell lung cancer (NSCLC): 181946 G/A (rs2293347), -191 C/A (rs712830) and -216G/T (rs712829). Materials and methods . The protocol for genotyping 181946C>T, 191C>A and -216G/T was refined according to previously reported data. Polymerase chain reaction (PCR) products measuring 197 bp were detected using electrophoresis in a 2% agarose gel, followed by staining with ethidium bromide. Results . The Biomaster HS Taq-PCR Color 2× and Biomaster LR HS PCR 2× reagent kits were effective for amplification of 181946 G/A polymorphism located in the intron of the EGFR gene. Additionally, polymorphisms -191 C/A (rs712829) and -216G/T (rs712829), located in the promoter region and containing a high GC content, were successfully amplified using the Biomaster LR HS PCR 2× kit. Conclusion . The present study shows that the Biomaster HS Taq-PCR Color 2× and Biomaster LR HS PCR 2× reagent kits are effective for amplification of 181946 G/A polymorphism located in the intron of the EGFR gene. Furthermore, the EGFR SNP -191 C/A, located in the promoter region with a high GC content, was successfully amplified using the Biomaster LR HS PCR 2× reagent kit.