A cohort of 45 clinically diagnosed Brugada Syndrome patients classified as SCN5A-negative was analyzed using next generation sequencing. Twenty-eight genes were resequenced. A total of 85 clinically evaluated relatives were also genetically analyzed to ascertain familial segregation. We identified genetic variants in novel candidate genes potentially associated to Brugada Syndrome. The identification of these genetic variants opens new perspectives on the implications of genetic background in the arrhythmogenic substrate for research purposes.