DNA transfection Mammalian cells - Immortalized cell lines OVCAR-3

Transfection is a powerful technique that enables the study of the function of genes and gene products in cells. Based on the nature of experiments, we may need a stable DNA transfection in cells for persistent gain-of-function or loss-of-function of the target gene. For stable transfection, integration of a DNA vector into the chromosome is crucial which requires selective screening and clonal isolation. By carefully selecting a viral delivery system and related reagents we can ensure safe and highly-efficient delivery of expression constructs for high-level constitutive or inducible expression in any mammalian cell type.

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Protocol tips
Plasmid transfections of 3×105 OVCAR3 cells seeded in 6-well plates were performed in antibiotic-free growth medium using FuGENE HD (Roche Diagnostic Corporation; Indianapolis, IN). Controls were treated with 100 ul/ml OPTI-MEM only and vehicle controls with 3ul/ml FuGENE HD
Protocol tips
The ovarian carcinoma cells were treated with RhoC-expressing plasmid by Attractene Transfection Reagent (QIAGEN) with pEGFP-N1 as a mock or RhoC siRNA (Sigma, USA) by HiPerFect Transfection Reagent (QIAGEN).
Protocol tips
Cells starved of glucose and/or glutamine for 24–36 hr were stimulated as indicated. Cells were transfected with JetPEI Polyplus reagent (Genycell Biotech) and treated as indicated 24 hr later.
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