{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T19:27:12Z","timestamp":1785353232681,"version":"3.55.0"},"reference-count":24,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2023,2,7]],"date-time":"2023-02-07T00:00:00Z","timestamp":1675728000000},"content-version":"vor","delay-in-days":6,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2023,2]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Duchenne Muscular Dystrophy (DMD) is an X\u2010linked inherited neuromuscular disorder caused by pathogenic variants in the dystrophin gene (<jats:italic>DMD<\/jats:italic>; locus Xp21.2). The variant spectrum of <jats:italic>DMD<\/jats:italic> is unique in that 65% of causative mutations are intragenic deletions, with intragenic duplications and point mutations (along with other sequence variants) accounting for 6% to 10% and 30% to 35%, respectively. The traditional strategy for molecular diagnostic testing for DMD involves initial screening for deletions\/duplications using microarray\u2010based comparative genomic hybridization followed by a full\u2010sequence analysis of <jats:italic>DMD<\/jats:italic> for sequence variants. This traditional strategy is expensive and time\u2010consuming due to the involvement of two separate tests to detect all types of variants in the <jats:italic>DMD<\/jats:italic> gene. Recent advancements in next\u2010generation sequencing (NGS) technology and improvements in analysis algorithms related to copy number variant detection ultimately resulted in the development of a single NGS\u2010based assay to detect all variant types, including deletions\/duplications and sequence variants. This article initially discusses the strategic algorithm for establishing a molecular diagnosis of DMD and later provides detailed molecular diagnostic protocols for DMD, including an NGS\u2010based sequencing assay with sequence and copy number variant analysis. \u00a9 2023 Wiley Periodicals LLC.<\/jats:p><jats:p><jats:bold>Basic Protocol<\/jats:bold>: Next\u2010generation sequencing of the entire genomic sequence of the <jats:italic>DMD<\/jats:italic> gene using IDT xGen Lockdown Probes<\/jats:p>","DOI":"10.1002\/cpz1.669","type":"journal-article","created":{"date-parts":[[2023,2,7]],"date-time":"2023-02-07T11:56:26Z","timestamp":1675770986000},"source":"Crossref","is-referenced-by-count":7,"title":["Molecular Diagnosis of Duchenne Muscular Dystrophy Using Single NGS\u2010Based Assay"],"prefix":"10.1002","volume":"3","author":[{"given":"Babi Ramesh Reddy","family":"Nallamilli","sequence":"first","affiliation":[{"name":"PerkinElmer Genomics PerkinElmer Inc  Waltham Massachusetts"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naga","family":"Guruju","sequence":"additional","affiliation":[{"name":"PerkinElmer Genomics PerkinElmer Inc  Waltham Massachusetts"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vanessa","family":"Jump","sequence":"additional","affiliation":[{"name":"PerkinElmer Genomics PerkinElmer Inc  Waltham Massachusetts"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruby","family":"Liu","sequence":"additional","affiliation":[{"name":"PerkinElmer Genomics PerkinElmer Inc  Waltham Massachusetts"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Madhuri","family":"Hegde","sequence":"additional","affiliation":[{"name":"PerkinElmer Genomics PerkinElmer Inc  Waltham 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