Rapid direct sequence analysis of the dystrophin gene

Kevin M. Flanigan, Andrew Von Niederhausern, Diane M. Dunn, Jonathan Alder, Jerry R. Mendell, Robert B. Weiss

Research output: Contribution to journalArticlepeer-review

147 Scopus citations

Abstract

Mutations in the dystrophin gene result in both Duchenne and Becker muscular dystrophy (DMD and BMD), as well as X-linked dilated cardiomyopathy. Mutational analysis is complicated by the large size of the gene, which consists of 79 exons and 8 promoters spread over 2.2 million base pairs of genomic DNA. Deletions of one or more exons account for 55%-65% of cases of DMD and BMD, and a multiplex polymerase chain reaction method - currently the most widely available method of mutational analysis - detects ∼98% of deletions. Detection of point mutations and small subexonic rearrangements has remained challenging. We report the development of a method that allows direct sequence analysis of the dystrophin gene in a rapid, accurate, and economical fashion. This same method, termed "SCAIP" (single condition amplification/internal primer) sequencing, is applicable to other genes and should allow the development of widely available assays for any number of large, multiexon genes.

Original languageEnglish (US)
Pages (from-to)931-939
Number of pages9
JournalAmerican journal of human genetics
Volume72
Issue number4
DOIs
StatePublished - Apr 1 2003
Externally publishedYes

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

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