title: Development of TaqMan allelic discrimination based genotyping of large DNA deletions.
abstract: The high prevalence of genetic diseases resulting from gross deletions has highlighted a need for a quick, simple, and reliable method of genotyping these mutations. Here, we developed a novel strategy for applying TaqMan allelic discrimination to accurately genotype 3 different large deletions in a high-throughput manner. Allelic discrimination has previously been used to genotype frame shift and point mutations, and small insertions or deletions six base pairs in length, but not large deletions. The assays designed here recognize a 2502 base pair deletion in the Nebulin (NEB) gene that results in Nemaline Myopathy, a 308,769 base pair deletion in the Gap Junction Protein, beta 6 (GJB6) gene that causes Hearing Loss, and a 6433 base pair deletion in the Mucolipin 1 (MCOLN1) gene responsible for causing Mucolipidosis IV Disease. This methodology may also be successfully applied to high throughput genotyping of other large deletions.
authors.affiliation: Department of Molecular Genetics, Microbiology, and Immunology, UMDNJ-Robert Wood Johnson Medical School, Piscataway, NJ, USA. afedick@rmanj.com
authors.forename: Anastasia
authors.initials: A
authors.lastname: Fedick
authors.forename: Jing
authors.initials: J
authors.lastname: Su
authors.forename: Nathan R
authors.initials: NR
authors.lastname: Treff
mesh_terms: D000483:Alleles; D001483:Base Sequence; D000075302:Connexin 30; D017630:Connexins / Q000235:genetics*; D060005:Genotyping Techniques / Q000379:methods*; D034381:Hearing Loss / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D009081:Mucolipidoses / Q000235:genetics; D009124:Muscle Proteins / Q000235:genetics*; D017696:Myopathies, Nemaline / Q000235:genetics; D020641:Polymorphism, Single Nucleotide; D017384:Sequence Deletion / Q000235:genetics*; D050051:Transient Receptor Potential Channels / Q000235:genetics*
chemical_list: D000075302:Connexin 30; D017630:Connexins; C491020:GJB6 protein, human; C416674:MCOLN1 protein, human; D009124:Muscle Proteins; D050051:Transient Receptor Potential Channels; C035620:nebulin