Grist S A, Firgaira F A, Morley A A
Flinders University of South Australia.
Biotechniques. 1993 Aug;15(2):304-9.
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. Using 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.
含有串联二核苷酸重复序列的DNA序列是人类和其他真核生物基因组中丰富的DNA多态性来源。在此，我们描述了一种用于鉴定和表征含有这些重复元件的DNA区域的新技术。使用设计用于识别串联二核苷酸重复序列的引物，并通过对目标基因组文库进行有限稀释，能够通过聚合酶链反应（PCR）扩增含有二核苷酸重复序列的单靶分子。扩增产物通过PCR直接法进行测序，并利用所得序列设计位点特异性引物。本研究鉴定并表征了四个匿名二核苷酸重复序列，其中三个表现出多态性。尽管该技术是针对二核苷酸重复序列开发的，但它普遍适用于通常通过PCR分析大小的重复DNA元件。该技术相对快速，省去了文库筛选及其相关操作，与现有的重复DNA回收方法相比具有优势。
