title: Single nucleotide polymorphism detection by polymerase chain reaction-restriction fragment length polymorphism.
abstract: Accurate analysis of DNA sequence variation in not only humans and animals but also other organisms has played a significant role in expanding our knowledge about genetic variety and diversity in a number of different biological areas. The search for an understanding of the causes of genetic variants and mutations has resulted in the development of a simple laboratory technique, known as the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method, for the detection of single nucleotide polymorphisms (SNPs). PCR-RFLP allows rapid detection of point mutations after the genomic sequences are amplified by PCR. The mutation is discriminated by digestion with specific restriction endonucleases and is identified by gel electrophoresis after staining with ethidium bromide (EtBr). This convenient and simple method is inexpensive and accurate for SNP genotyping and especially useful in small basic research studies of complex genetic diseases. The whole protocol takes only a day to carry out.
authors.affiliation: Department of Legal Medicine, Shinshu University School of Medicine, Nagano 390-8621, Japan. otamasao@sch.md.shinshu-u.ac.jp
authors.forename: Masao
authors.initials: M
authors.lastname: Ota
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Fukushima
authors.forename: Jerzy K
authors.initials: JK
authors.lastname: Kulski
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Inoko
mesh_terms: D042942:3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide) / Q000235:genetics; D000017:ABO Blood-Group System / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D004262:DNA Restriction Enzymes; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005838:Genotype; D006684:HLA-DR Antigens / Q000737:chemistry / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide
chemical_list: D000017:ABO Blood-Group System; D006684:HLA-DR Antigens; D042942:3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide); D004262:DNA Restriction Enzymes