Dual report assay system

Manufactured by Promega

The Dual-Luciferase® Reporter Assay System is a sensitive and versatile tool for gene expression analysis. It provides a method for quantifying two reporter enzyme activities within a single sample. The system uses the firefly and Renilla luciferase enzymes as reporter proteins.

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Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions) Pgl4.17 vector, by Promega (1 mentions)

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Dual-Luciferase Report Assay System

2 protocols using dual report assay system

Zebrafish Promoter Luciferase Assay

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A 2,129-bp zebrafish cebpb promoter fragment or a 1,999-bp zebrafish ulk1a promoter fragment was PCR amplified and cloned into pGL4.17 vector (Promega, 9PIE672), respectively. Human embryonic kindey (HEK) 293T cells were cultured in DMEM containing 10% serum and penicillin-streptomycin in a 24-well plate. Transfection was done with Lipofectamine 2000 (Invitrogen, 11668027) according the manufacturer's instructions. Luciferase reporter assays were performed with a Dual-Report assay system (Promega, E1980). 100 ng each of cebpb-luc, ulk1a-luc, clocka/clock1a, arntl1b/bmal1b and cry1ab were used; for the dosage-dependent experiment, 100, 200 and 300 ng of nr1d1 or roraa, were used. The total DNA amounts were equalized by pcDNA 3.1 vector. Three independent experiments were conducted for each assay.

Huang G., Zhang F., Ye Q, & Wang H. (2016). The circadian clock regulates autophagy directly through the nuclear hormone receptor Nr1d1/Rev-erbα and indirectly via Cebpb/(C/ebpβ) in zebrafish. Autophagy, 12(8), 1292-1309.

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Transfection Assay in SCN2.2 Cells

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SCN2.2 cells (RRID:CVCL_D050) were cultured in MEM containing 10% FBS, 0.4% glucose, and 2 mM L-glutamine in 96-well dishes (Earnest et al., 1999 (link)). Transfection assay was performed by using Lipofectamine 2000 (Thermo Fisher Scientific) according to the manufacturer’s instructions. Luciferase reporter gene assays were carried out with a Dual-Report assay system (Promega); 50 ng Prokr2-Luc, 50 ng ZBTB20, 50 ng P300, 1 ng Renilla pRL-TKvector were added and the pCMV-Tag2B plasmid was brought to the same amount.

Qu Z., Zhang H., Huang M., Shi G., Liu Z., Xie P., Li H., Wang W., Xu G., Zhang Y., Yang L., Huang G., Takahashi J.S., Zhang W.J, & Xu Y. (2016). Loss of ZBTB20 impairs circadian output and leads to unimodal behavioral rhythms. eLife, 5, e17171.

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