Experiment Tags: Lentiviral Vectors, Stable Cell Line, Gene Expression, Viral Transduction, Cell Culture, Puromycin Selection, Virus Packaging, Mammalian Cells, Transgene Expression
Experiment Summary: This protocol details the generation of stable cell lines using lentiviral vectors. The method covers plasmid preparation, 293T cell transfection for viral packaging, PEG8000-based virus concentration, target cell infection, antibiotic selection parameters, and single-clone isolation. Includes MOI calculation guidance, troubleshooting for common challenges like low viral titer and poor infection efficiency, and validation strategies for ensuring stable transgene expression.