Language
English
Publication Date
1-1-2024
Journal
Methods in Molecular Biology
DOI
10.1007/978-1-0716-3770-8_6
PMID
38814393
Abstract
Here we describe the in vitro preparation of mRNA from DNA templates, including setting up the transcription reaction, mRNA capping, and mRNA labeling. We then describe methods used for mRNA characterization, including UV and fluorescence spectrophotometry, as well as gel electrophoresis. Moreover, characterization of the in vitro transcribed RNA using the Bioanalyzer instrument is described, allowing a higher resolution analysis of the target molecules. For the in vitro testing of the mRNA molecules, we include protocols for the transfection of various primary cell cultures and the confirmation of translation by intracellular staining and western blotting.
Keywords
RNA, Messenger, Transcription, Genetic, Humans, Transfection, RNA Caps, DNA, Animals, Flow cytometry, Fluorescence spectrophotometry, Immunofluorescence microscopy, In vitro transcription, Intracellular staining, LNPs, Lipid nanoparticles, Ultraviolet spectrophotometry, Western blotting, mRNA capping
Published Open-Access
yes
Recommended Citation
Cristina Poveda, Yi-Lin Chen, and Ulrich Strych, "Generation and Characterization of In Vitro Transcribed mRNA" (2024). Faculty, Staff and Students Publications. 6491.
https://digitalcommons.library.tmc.edu/baylor_docs/6491