In Vitro RNA Transcription and Electroporation

After linearization of the plasmids described in the former paragraph using XbaI and treatment with the Mung Bean Nuclease (New England Biolabs, Ipswich, MA, USA) as previously described [8 (link)], RNA was in vitro transcribed using MEGAscript^® T7 transcription kit (Life Technologies, Carlsbad, CA, USA). Huh-7 cells were electroporated using 10 μg of RNA according to the method previously described [51 (link)], using 0.2 cm gap electroporation cuvettes (Biorad, Hercules, CA, USA).

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Riva L., Spriet C., Barois N., Popescu C.I., Dubuisson J, & Rouillé Y. (2021). Comparative Analysis of Hepatitis C Virus NS5A Dynamics and Localization in Assembly-Deficient Mutants. Pathogens, 10(2), 172.

Publication 2021

Mung Bean Nuclease MEGAscript® T7 transcription kit

Cells Electroporation Mung bean nuclease Plasmids Transcription

Corresponding Organization :

Other organizations : Center for Infection and Immunity of Lille, Institut Pasteur de Lille, Centre National de la Recherche Scientifique, Centre Hospitalier Universitaire de Lille, Université de Lille, Inserm, Unité de Glycobiologie Structurale et Fonctionnelle

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Variable analysis

independent variables

Linearization of the plasmids using XbaI
Treatment with the Mung Bean Nuclease (New England Biolabs, Ipswich, MA, USA)

dependent variables

RNA in vitro transcription using MEGAscript® T7 transcription kit (Life Technologies, Carlsbad, CA, USA)
Electroporation of Huh-7 cells with 10 μg of RNA

control variables

Use of 0.2 cm gap electroporation cuvettes (Biorad, Hercules, CA, USA)

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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