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An Efficient Electroporation Protocol for the Genetic Modification of Mammalian Cells.

Front Bioeng Biotechnol. 2016; 
Chicaybam Leonardo,Barcelos Camila,Peixoto Barbara,Carneiro Mayra,Limia Cintia Gomez,Redondo Patrícia,Lira Carla,Paraguassú-Braga Flávio,Vasconcelos Zilton Farias Meira De,Barros Luciana,Bonamino Martin Her
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Gene Synthesis /NotI, and the neomycin resistance gene (NEO), which was synthesized by Genscript (Piscataway, NJ Get A Quote

摘要

Genetic modification of cell lines and primary cells is an expensive and cumbersome approach, often involving the use of viral vectors. Electroporation using square-wave generating devices, like Lonza's Nucleofector, is a widely used option, but the costs associated with the acquisition of electroporation kits and the transient transgene expression might hamper the utility of this methodology. In the present work, we show that our in-house developed buffers, termed Chicabuffers, can be efficiently used to electroporate cell lines and primary cells from murine and human origin. Using the Nucleofector II device, we electroporated 14 different cell lines and also primary cells, like mesenchymal s... More

关键词

CD34,CRISPR,GFP,MSC,PD-1,T lymphocyte,cell line,electroporation,transp