Dinucleotide repeat polymorphisms isolated by the polymerase chain reaction

S. A. Grist, F. A. Firgaira, A. A. Morley

    Research output: Contribution to journalArticle

    28 Citations (Scopus)

    Abstract

    DNA sequences containing tandem dinucleotide repeats represent an abundant source of DNA polymorphism in human and other eukaryotic genomes. Here we describe a novel technique for the identification and characterization of regions of DNA containing these repetitive elements. Usng primers designed to recognize tandem dinucleotide repeat sequences and limiting dilution of a target genomic library enable amplification by polymerase chain reaction (PCR) of single-target molecules containing dinucleotide repeats. Amplified material was sequenced by the PCR direct method and by the resultant sequences used to design locus-specific primers. This study identified and characterized four anonymous dinucleotide repeat sequences, three of which exhibited polymorphism. Although developed for dinucleotide repeats, the technique is universally applicable to repeat DNA elements of a size usually analyzed by PCR. The technique is comparatively rapid, eliminates library screening and its associated manipulations, and compares favorably with existing methods for the recovery of repetitive DNAs.

    Original languageEnglish
    Pages (from-to)304-309
    Number of pages6
    JournalBiotechniques
    Volume15
    Issue number2
    Publication statusPublished - 1 Aug 1993

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