In ovo electroporation is a well-established method of gene transfer into neural and mesenchymal tissue in chick embryos. Electroporation of somites, however, has been hampered by low efficiency due to technical difficulties. Here, we present a powerful technique to electroporate avian somites and subpopulations of somitic cells at high efficiency in ovo. We demonstrate specific targeting of distinct somitic compartments and their derivatives using single or combinations of plasmid expression vectors. This technique opens new perspectives to investigate the morphologic and genetic basis of somite development. Developmental Dynamics 229:643–650, 2004. © 2004 Wiley-Liss, Inc.