title: Tetra Primer ARMS PCR Optimization to Detect Single Nucleotide Polymorphisms of the CYP2E1 Gene.
abstract: Single nucleotide polymorphism (SNP) detection has been used extensively for genetic association studies of diseases including cancer. For mass, yet accurate and more economic SNP detection we have optimized tetra primer amplification refractory mutation system polymerase chain reaction (ARMS PCR) to detect three SNPs in the cytochrome P450 2E1 (CYP2E1) gene locus; i.e. rs3813865, rs2070672 and rs3813867. The optimization system strategies used were (1) designing inner and outer primers; (2) determining of their optimum primer concentration ratios; and (3) determining of the optimum PCR annealing temperature. The tetra primer ARMS PCR result could be directly observed using agarose gel electrophoresis. The method succesfully determined three SNPs in CYP2E1 locus, the results being consistent with validation using DNA sequencing and restriction fragment length polymorphisms (RFLP).
authors: Biomedical Science, Faculty of Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia Email : jajahfachiroh@ugm.ac.id.
authors: Saihas
authors: S
authors: Suhda
authors: Dewi Kartikawati
authors: DK
authors: Paramita
authors: Jajah
authors: J
authors: Fachiroh
mesh_terms: D000818:Animals; D060126:Characidae / Q000235:genetics*; D019392:Cytochrome P-450 CYP2E1 / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics*; D004587:Electrophoresis, Agar Gel / Q000379:methods; D009154:Mutation / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length / Q000235:genetics; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*
chemical_list: D017931:DNA Primers; D019392:Cytochrome P-450 CYP2E1