Cloning and polymerase chain reaction-single-strand conformation polymorphism analysis of anonymous Alu repeats on chromosome 11

M Iizuka; S Mashiyama; M Oshimura; T Sekiya; K Hayashi

doi:10.1016/0888-7543(92)90416-p

Cloning and polymerase chain reaction-single-strand conformation polymorphism analysis of anonymous Alu repeats on chromosome 11

Genomics. 1992 Jan;12(1):139-46. doi: 10.1016/0888-7543(92)90416-p.

Authors

M Iizuka¹, S Mashiyama, M Oshimura, T Sekiya, K Hayashi

Affiliation

¹ Oncogene Division, National Cancer Center Research Institute, Tokyo, Japan.

PMID: 1733851
DOI: 10.1016/0888-7543(92)90416-p

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacteriophages / genetics
Base Sequence
Chromosomes, Human, Pair 11*
Cloning, Molecular
DNA, Single-Stranded / chemistry*
Genetic Vectors
Genomic Library
Humans
Molecular Sequence Data
Nucleic Acid Conformation
Plasmids
Polymerase Chain Reaction / methods*
Polymorphism, Genetic*
Repetitive Sequences, Nucleic Acid*

Substances

DNA, Single-Stranded