title: Development of an automated SNP analysis method using a paramagnetic beads handling robot.
abstract: Biological and medical importance of the single nucleotide polymorphism (SNP) has led to development of a wide variety of methods for SNP typing. Aiming for establishing highly reliable and fully automated SNP typing, we have developed the adapter ligation method in combination with the paramagnetic beads handling technology, Magtration(R). The method utilizes sequence specific ligation between the fluorescently labeled adapter and the sample DNAs at the cohesive end produced by a type IIS restriction enzyme. Evaluation of the method using human genomic DNA showed clear discrimination of the three genotypes without ambiguity using the same reaction condition for any SNPs examined. The operations following PCR amplification were automatically performed by the Magtration(R)-based robot that we have previously developed. Multiplex typing of two SNPs in a single reaction by using four fluorescent dyes was successfully preformed at the almost same sensitivity and reliability as the single typing. These results demonstrate that the automated paramagnetic beads handling technology, Magtration(R), is highly adaptable to the automated SNP analysis and that our method best fits to an automated in-house SNP typing for laboratory and medical uses.
authors.affiliation: Advanced Institute of Industrial Science and Technology (AIST), Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Hagiwara
authors.forename: Kazumi
authors.initials: K
authors.lastname: Sawakami-Kobayashi
authors.forename: Midori
authors.initials: M
authors.lastname: Yamamoto
authors.forename: Shoji
authors.initials: S
authors.lastname: Iwasaki
authors.forename: Mika
authors.initials: M
authors.lastname: Sugiura
authors.forename: Hatsumi
authors.initials: H
authors.lastname: Abe
authors.forename: Sumiko
authors.initials: S
authors.lastname: Kunihiro-Ohashi
authors.forename: Kumiko
authors.initials: K
authors.lastname: Takase
authors.forename: Noriko
authors.initials: N
authors.lastname: Yamane
authors.forename: Kaoru
authors.initials: K
authors.lastname: Kato
authors.forename: Renkon
authors.initials: R
authors.lastname: Son
authors.forename: Michihiro
authors.initials: M
authors.lastname: Nakamura
authors.forename: Osamu
authors.initials: O
authors.lastname: Segawa
authors.forename: Mamiko
authors.initials: M
authors.lastname: Yoshida
authors.forename: Masafumi
authors.initials: M
authors.lastname: Yohda
authors.forename: Hideji
authors.initials: H
authors.lastname: Tajima
authors.forename: Masato
authors.initials: M
authors.lastname: Kobori
authors.forename: Yousuke
authors.initials: Y
authors.lastname: Takahama
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Itakura
authors.forename: Masayuki
authors.initials: M
authors.lastname: Machida
mesh_terms: D004867:Equipment Design / Q000295:instrumentation; D005820:Genetic Testing; D015894:Genome, Human; D005838:Genotype; D006801:Humans; D008280:Magnetics; D016133:Polymerase Chain Reaction / Q000295:instrumentation*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D012371:Robotics / Q000295:instrumentation*; D017422:Sequence Analysis, DNA / Q000295:instrumentation*