GenJet™ In Vitro DNA Transfection Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- For high efficiency, transfect cells at high density. 70~80% confluency is highly recommended - To lower cytotoxicity, transfect cells in presence of serum (10%) and antibiotics

Upstream tips
- For high efficiency, transfect cells at high density. 70~80% confluency is highly recommended - To lower cytotoxicity, transfect cells in presence of serum (10%) and antibiotics

Publication protocol

Transient and siRNA transfections
MCF-7 cells were cultured for 18 hrs before transient transfection with each plasmid using GenJet™ Reagent II for MCF-7 (Signagen, Ijamsville, MD) according to the manufacturer's instructions. The plasmid expression was confirmed through changes in cell shape, and immunoblot or fluorescence analyses. The following plasmids (Addgene, Cambridge, MA) were used: pCDNA3-EGFP (Addgene ID 13031), pCDNA3-EGFP-RhoA-Q63L (constitutively active RhoA, Addgene ID 12968), pCDNA3-EGFP-RhoA-T19N (dominant negative, Addgene ID 12967), pCDNA3-EGFP-Rac1-Q61L (constitutively active Rac1, Addgene ID 12981), pCDNA3-EGFP-Rac1-T17N (dominant negative, Addgene ID 12982), and pCDNA3-GFP-PTEN (wild type, Addgene ID 10759). For the RNA interference (RNAi) experiments, MCF-7 cells were seeded for 18 hrs and transfected with siRNA (Santa Cruz Biotechnology, Santa Cruz, CA) using a siRNA transfection reagent (Santa Cruz Biotechnology, Santa Cruz, CA) according to manufacturer's instructions. The siRNA for targeting ROCK-1, ROCK-2, and PTEN were obtained from Santa Cruz Biotechnology (Santa Cruz Biotechnology, Santa Cruz, CA).



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Manufacturer protocol

Download the product protocol from SignaGen Laboratories for GenJet™ In Vitro DNA Transfection Reagent below.

Download manufacturer protocol

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