Detection of single nucleotide polymorphism in the FUT2 gene by temperature gradient gel electrophoresis.
A simple and rapid analysis system for single nucleotide polymorphisms (SNPs) was investigated for the FUT2 gene using the temperature gradient gel electrophoresis (TGGE) method. The 426-bp or 259-bp FUT2 fragments were amplified from heterozygous samples using primers, and the heteroduplex and homoduplex bands were detected by TGGE. The FUT2 fragments amplified from homozygous samples were denatured and re-annealed with a known sequence fragment, forming heteroduplex bands which were analyzed by TGGE. The fragment patterns of homoduplex and heteroduplex bands in TGGE were specific to the genotypes, and more specifically, the number of heteroduplex bands and the migration of the homoduplex bands corresponded to the number of nucleotide substitutions and the change in GC content due to the substitutions, respectively. Thus, the TGGE technique can act as a high-throughput method for the detection and the presumption of sequences of known and unknown SNPs in the FUT2 gene.
Mitani
Tomoaki
T
Department of Legal Medicine, Kansai Medical University, Japan; Forensic Science Laboratory, Osaka Prefectural Police Headquarters, Honmachi 1-3-18, Chuo-ku, Osaka 541-0053, Japan. mitani@bea.hi-ho.ne.jp
Akane
Atsushi
A
D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D005647:Fucosyltransferases / Q000235:genetics*; D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D017421:Sequence Analysis; D000097631:Galactoside 2-alpha-L-fucosyltransferase
D017931:DNA Primers; D005647:Fucosyltransferases; D000097631:Galactoside 2-alpha-L-fucosyltransferase