Multiplex Ligation-dependent Probe Amplification (MLPA)
MLPA kit for DMD (Salsas PO34-PO35) was used to find deletions/duplications (Gatta et al., 2005; Janssen et al., 2005; Schwartz & Dunø, 2004). Reactions were performed according to the manufacturer’s recommendations [MRC-Holland, Amsterdam, The Netherlands (www.mlpa.com)]. Products were analyzed using a fragment analyzer sequencer [ABI 3730XL; Applied Biosystems, Foster City, California] and 500Liz as internal size standard. Data analysis was performed using Coffalyser [MRC-Holland, Amsterdam, The Netherlands] and GeneMarker V2.2.0 [Softgenetics, State College, Pennsylvania] softwares. Wildtype, deleted, and duplicated controls were included.