In Vitro Luciferase Assay Protocol

In vitro translation assays were performed as previously described [15 (link)]. 3 pmol of RNA transcripts were used in a 25 μL translation reaction using wheat germ extract (WGE) (Promega) according to the manufacturer′s instructions. The luciferase activity was measured using a luciferase assay reporter system (Promega) and a Modulus Microplate Multimode Reader (Turner BioSystems). At least three independent in vitro translationa ssays were performed for each construct. Standard errors were calculated in Microsoft Excel.

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Wang D., Yang C., Deng Y., Cao X., Xu W., Han Z., Li Q., Yang Y, & Yuan X. (2022). Conserved RNA secondary structure in Cherry virus A 5′-UTR associated with translation regulation. Virology Journal, 19, 91.

Publication 2022

luciferase assay reporter system Modulus Microplate Multimode Reader

Assay Luciferase Promega Wheat

Corresponding Organization : Zaozhuang University

Other organizations : Shandong Agricultural University

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Protocol cited in 1 other protocol

Variable analysis

independent variables

RNA transcripts

dependent variables

Luciferase activity

control variables

Translation reaction volume (25 μL)
Wheat germ extract (WGE) (Promega)
Luciferase assay reporter system (Promega)
Modulus Microplate Multimode Reader (Turner BioSystems)

controls

Positive control: Wheat germ extract (WGE) (Promega) used for in vitro translation assays
Negative control: Not explicitly mentioned

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