MELOE‐1 intercistronic region IR1215‐1490 (275 nt IRES region) [20 (link)] was cloned into a pBSSK vector under the control of a T7 promoter. It was linearized with NotI and in vitro transcribed according to the MegaShort Script Kit (Ambion, Thermo Fisher Scientific) protocol. RNA (0.2 nmol) was biotinylated using the 5’end modification kit and biotin maleimide (Vector Laboratories, Eurobio Scientific, Les Ulis, France) following the manufacturer’s guidelines.
Charpentier M., Dupré E., Fortun A., Briand F., Maillasson M., Com E., Pineau C., Labarrière N., Rabu C, & Lang F. (2021). hnRNP‐A1 binds to the IRES of MELOE‐1 antigen to promote MELOE‐1 translation in stressed melanoma cells. Molecular Oncology, 16(3), 594-606.
Other organizations :
Inserm, Nantes Université, Centre National de la Recherche Scientifique, Structure Fédérative de Recherche en Biologie et Santé de Rennes, École des Hautes Études en Santé Publique, Université de Rennes, Institut de Recherche en Santé, Environnement et Travail
Biotinylation of 0.2 nmol of RNA using 5'end modification kit and biotin maleimide
control variables
Incubation with manufacturer's guidelines for biotinylation
controls
Positive control: None mentioned
Negative control: None mentioned
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