Analyzing Copy Number Variation Using Pulsed-Field Gel Electrophoresis: Providing a Genetic Diagnosis for FSHD1.
The myopathy facioscapulohumeral muscular dystrophy type 1 (FSHD1) is caused by copy number variation of the D4Z4 macrosatellite repeat on chromosome 4. In unaffected individuals the number of 3.3 kb D4Z4 units varies between 8 and 100, whereas 1-10 units are seen in FSHD1 cases. A homologous and heterogenous D4Z4 array can be found on chromosome 10q, but contractions of this array are typically not associated with FSHD. Discriminating between the chromosome 4 and chromosome 10 D4Z4 arrays, as well as determining the array size, requires the use of pulsed-field gel electrophoresis, Southern blotting, and the isolation of high-quality DNA.
Department of Human Genetics, Leiden University Medical Center, 2333, ZA, Leiden, The Netherlands. R.J.L.F.Lemmers@lumc.nl.
Richard J L F
RJ
Lemmers
D002879:Chromosomes, Human, Pair 10; D056915:DNA Copy Number Variations; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods*; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D025202:Molecular Diagnostic Techniques; D020391:Muscular Dystrophy, Facioscapulohumeral / Q000175:diagnosis / Q000235:genetics*
C121257:DUX4L1 protein, human; D018398:Homeodomain Proteins
Copy number variation (CNV); D4Z4; DNA agarose plug; FSHD; Macrosatellite repeat; Pulsed-field gel electrophoresis (PFGE); Southern blot