title: SNP analysis of the inter-alpha-trypsin inhibitor family heavy chain-related protein (IHRP) gene by a fluorescence-adapted SSCP method.
abstract: BACKGROUND
  Single-nucleotide polymorphisms (SNPs) are considered to be useful polymorphic markers for genetic studies of polygenic traits. Single-stranded conformational polymorphism (SSCP) analysis has been widely applied to detect SNPs, including point mutations in cancer and congenital diseases. In this study, we describe an application of the fluorescent labeling of PCR fragments using a fluorescent-adapted primer for SSCP analysis as a novel method.
  
  
  METHODS
  Single-nucleotide polymorphisms (SNPs) of the inter-alpha-trypsin inhibitor family heavy chain-related protein (IHRP) gene were analyzed using a fluorescence-adapted SSCP method. The method was constructed from two procedures: 1) a fluorescent labeling reaction of PCR fragments using fluorescence-adapted primers in a single tube, and 2) electrophoresis on a non-denaturing polyacrylamide gel.
  
  
  RESULTS
  This method was more economical and convenient than the single-stranded conformational polymorphism (SSCP) methods previously reported in the detection of the labeled fragments obtained. In this study, eight SNPs of the IHRP gene were detected by the fluorescence-adapted SSCP. One of the SNPs was a new SNP resulting in an amino acid substitution, while the other SNPs have already been reported in the public databases. Six SNPs of the IHRP were associated with two haplotypes.
  
  
  CONCLUSIONS
  The fluorescence-adapted SSCP was useful for detecting and genotyping SNPs.
authors.affiliation: Department of Physiological Chemistry, School of Pharmaceutical Sciences, Showa University, 1-5-8 Hatanodai, Shinagawa, Tokyo 142-8555, Japan. ttozaki@nyc.odn.ne.jp
authors.forename: Teruaki
authors.initials: T
authors.lastname: Tozaki
authors.forename: Nam-Ho
authors.initials: NH
authors.lastname: Choi-Miura
authors.forename: Matsuo
authors.initials: M
authors.lastname: Taniyama
authors.forename: Masahiko
authors.initials: M
authors.lastname: Kurosawa
authors.forename: Motowo
authors.initials: M
authors.lastname: Tomita