title: Cloning and polymerase chain reaction-single-strand conformation polymorphism analysis of anonymous Alu repeats on chromosome 11.
abstract: We have shown that many of the Alu repeats found in the GenBank database are polymorphic and that this polymorphism can be detected by a simple technique, single-strand conformation polymorphism (SSCP) analysis, after polymerase chain reaction (PCR) amplification of each repeat from DNA of individuals. Here, we describe a method for collecting many anonymous Alu repeats and their flanks in a chromosome-specific phage library and cloning them into plasmids. The flanking single-copy sequences of each repeat in the plasmid were then determined, and 20mer to 30mer segments of these sequences were used as primers for the PCR-SSCP analysis. Many new polymorphic DNA markers on chromosome 11 were obtained with this method. These markers can also serve as sequence-tagged sites for physical mapping of the genome.
authors.affiliation: Oncogene Division, National Cancer Center Research Institute, Tokyo, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Iizuka
authors.forename: S
authors.initials: S
authors.lastname: Mashiyama
authors.forename: M
authors.initials: M
authors.lastname: Oshimura
authors.forename: T
authors.initials: T
authors.lastname: Sekiya
authors.forename: K
authors.initials: K
authors.lastname: Hayashi
mesh_terms: D001435:Bacteriophages / Q000235:genetics; D001483:Base Sequence; D002880:Chromosomes, Human, Pair 11; D003001:Cloning, Molecular; D004277:DNA, Single-Stranded / Q000737:chemistry*; D005822:Genetic Vectors; D015698:Genomic Library; D006801:Humans; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D012091:Repetitive Sequences, Nucleic Acid
chemical_list: D004277:DNA, Single-Stranded