title: Application of the RLGS method to large-size genomes using a restriction trapper.
abstract: We developed a method for producing restriction landmark genomic scanning (RLGS) profiles of large-size genomes, such as those of higher plants or amphibians using a restriction trapper. Use of the conventional RLGS method is limited to genomes smaller than 3 x 10(9) bp, because the larger genomic DNAs, especially those of more than 1 x 10(10) bp, produce high background due to incorporation of radioactivity at non-specifically damaged sites. Our new method reduces the background levels by reducing genome complexity to 1/200-1/300 using a purification step to enrich DNA fragments carrying specific restriction landmarks at their ends using a restriction trapper. This step makes it possible to obtain RLGS patterns of larger genomes. Our paper describes the practical application for the RLGS method using a restriction trapper with the pine tree genome (3 x 10(10) bp/haploid genome; Pinus koraiensis Sieb. et Zucc.) as an example.
authors.affiliation: Gene Bank, RIKEN Tsukuba Life Science Center, Ibaraki, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Okuizumi
authors.forename: Y
authors.initials: Y
authors.lastname: Okazaki
authors.forename: N
authors.initials: N
authors.lastname: Sasaki
authors.forename: M
authors.initials: M
authors.lastname: Muramatsu
authors.forename: K
authors.initials: K
authors.lastname: Nakashima
authors.forename: K
authors.initials: K
authors.lastname: Fan
authors.forename: H
authors.initials: H
authors.lastname: Tano
authors.forename: K
authors.initials: K
authors.lastname: Ohba
authors.forename: Y
authors.initials: Y
authors.lastname: Hayashizaki
mesh_terms: D018744:DNA, Plant / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D015183:Restriction Mapping; D014197:Trees / Q000235:genetics*
chemical_list: D018744:DNA, Plant