Typing Dinucleotide Repeats Under Nondenaturing Conditions with Single-Base Resolution and High Sizing Precision

ISSN： 1073-6085

Source： Molecular Biotechnology, Vol.21, Iss.2, 2002-06, pp. : 117-122

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Abstract

Dinucleotide repeats are genetic markers that are useful for many purposes, including genetic epidemiology, population genetics, and genetic diagnostics. The accuracy of analyses based on dinucleotide repeat polymorphisms is highly dependent on the success achieved in minimizing genotyping errors. Genotyping errors in dinucleotide repeat typing may arise for various reasons, including polymerase chain reaction (PCR) processing errors and the use of unsuitable electrophoretic conditions for resolving amplification products (i.e., lack of single-base resolution and inadequate precision in allele sizing). We have recently described a nondenaturing electrophoretic system useful for detecting PCR processing errors that lead to misidentification of heterozygotes as homozygotes in (AC)_n repeat typing. Here, we show that this system also allows resolution of (AC)_n repeats in native conditions with single-base resolution and high sizing precision, on the basis of an analysis of seven human (AC)_n repeats ranging in size from 72 to 217 bp. This PAGE system is thus also useful for reducing the likelihood both of allele misidentification due to the absence of single-base resolution and of inaccuracies in allele sizing due to anomalous electrophoretic migrations among the alleles within an (AC)_nrepeat.