Transfection describes the introduction of nucleic acid (DNA and RNA) to eukaryotic cells using non-viral methods. However, almost all of the classical transfection methods including electroporation exhibit a limited ability to transfect certain cell types, such as primary, neuronal, differentiated, and non-dividing cells. Nucleofection technology, marked by Lonza (Amaxa), though an electroporation-based method, is commonly used to effectively deliver siRNA and plasmids into these historically recalcitrant eukaryotic cell types because of its unique feature of directly delivering nucleic acid into nuclei. Additionally, the use of the nucleofection technique allows efficient transfection of stem cells with reduced risk of modifying cellular processes. The nucleofection technology requires the use of the Amaxa Nucleofector, combined with a set of special transfection buffers that greatly improves the transfection efficiency and increases the cell survival rate. The use of proprietary Nucleofection buffers which must be purchased directly from the vendor (Lonza) presents following limitations on the use of such a technology. Firstly, the nucleofection solutions are very costly for a small quantity preventing its routine use for transfections. Secondly, nucleofection buffers vary from cell type/line to cell type/line, and only limited cell types have an optimal solution available. Striving to become the world’s leading enterprise for mammalian genome engineering, GenomeFrontier, Biosciences, Inc. devoted one of her major efforts in establishing non-viral mammalian genome manipulating platform consisting of novel piggyBac transposon-based vector systems and buffer systems for nucleofection. Consequently, the universal, highly efficient, and cost-effective “ALL-IN-ONE” nucleofection buffer system is successfully developed and became commercially available.