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

β-Catenin/TCF Transcription Reporter Assay
HOBs plated in 24-well plates at 2 × 104/cm2 were transiently transfected with the Wnt-luciferase reporter construct pBAR (1µg total; Dr. RT. Moon, University of Washington, Seattle, WA, USA) and the control reporter pfuBAR (1µg total; Dr. RT. Moon, University of Washington, Seattle, WA, USA) using GeneJuice (Novagen, Madison, WI, USA), as previously reported20. The β-catenin activated reporter (pBAR) used to transfect HOBs contains 12 TCF binding sites (5’-AGATCAAAGG-3’) separated by distinct 5 base linkers. These elements lie directly upstream of a minimal thymidine kinase promoter driving firefly luciferase expression. The control reporter pfuBAR is identical to its sister reporter, except for a two base substitution in each TCF binding site. Both reporters contain a separate PGK promoter constitutively driving the expression of a puromycin resistance gene, and both are in a lentiviral platform. Subsequent co-transfection with 0.5µg of pSL9EF1a(P)RLUC (Dr. RT. Moon, University of Washington, Seattle, WA, USA), the internal control reporter driving constitutive expression of Renilla luciferase, was performed, normalizing for transfection efficiency. Sixteen hours after transfection, HOBs were washed and cultured for 48h in media containing gp120 with 2% FCS, in the presence or absence Wnt3a-conditioned media. HOBs were lysated and luciferase assays performed with the Dual Luciferase Assay Kit (Promega, Madison, WI, USA) as per manufacturer’s protocols and as previously reported20. 10µl of cell lysates were first assayed for firefly luciferase and then Renilla luciferase activity. Firefly luciferase activity was subsequently normalized to Renilla luciferase activity.

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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

Videos

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