GeneJuice® Transfection Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Use only high-quality DNA. If necessary, include an endotoxin removal step.	- GeneJuice®Transfection Reagent is compatible with both serum-containing and serum-free media. - But serum must not be present during formation of the GeneJuice Transfection Reagent/DNA complex.

Upstream tips
Use only high-quality DNA. If necessary, include an endotoxin removal step.
Protocol tips
- GeneJuice®Transfection Reagent is compatible with both serum-containing and serum-free media. - But serum must not be present during formation of the GeneJuice Transfection Reagent/DNA complex.

Publication protocol

siRNA, plasmid transfection and Cell culture
Mouse 3T3-L1 preadipocytes, mouse macrophage RAW264.7 cells and human HEK293 cells were cultured in a maintenance medium (10% fetal bovine serum, 100 units/ml penicillin and 100 μg/ml streptomycin in Dulbecco’s modified medium) at 37° in 5% CO2/95% humidified air. 3T3-L1 cells were differentiated as previously described20,22. Stimulation of RAW264.7 cells and co-cultivation of 3T3-L1 cells and RAW264.7 cells were performed based on previous protocol20,22. Differentiated 3T3-L1 adipocytes were stimulated with 10 ng/ml of mouse recombinant tumor necrosis factor-α (R&D system, Minneapolis, MN) for 24 hr. cDNA was amplified using a PCR primer set (5′-CCGCGTTCATGCACCGCCTG-3′ and 5′-ACCCGCGCCGCGCTAGCAGT-3′) designed according to the nucleotide sequences of mouse C/EBPβ and subcloned into pcDNA3.1, which generated pcDNA-C/EBPβ. DNA transfections were performed using GeneJuice Transfection Reagent (Merck Millipore) according to the manufacturer’s instructions. Small interfering RNA (siRNA) duplex oligoribonucleotides against mouse C/EBPβ were synthesized by Sigma.

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Papers

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Manufacturer protocol

Download the product protocol from Millipore for GeneJuice® Transfection Reagent below.

Download manufacturer protocol

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