Methylation Analysis using Sequenom MassARRAY

For methylation analysis, the Sequenom MassARRAY EpiTYPER Assay was applied as described previously [61 (link)]. The PCR amplicons were conducted subsequently according to the protocol of the Sequenom EpiTYPER Assay and cleaned by Resin. A nanodispenser was used to transfer the products to a 384 SpectroCHIP (SEQUENOM, San Diego, CA, USA). The chips were read by a Sequenom Mass Spectrometer system (SEQUENOM, San Diego, CA, USA). Data was gathered by SpectroACQUIRE v3.3.1.3 software (SEQUENOM, San Diego, CA, USA) and visualized with MassArray EpiTYPER v1.0 software (SEQUENOM, San Diego, CA, USA). Results were depicted as “beta” values (β) between 0 and 1.