Isolation of Newly Transcribed RNA Using the Metabolic Label 4-Thiouridine.
- Author(s): Garibaldi, Angela
- Carranza, Francisco
- Hertel, Klemens J
- et al.
Published Web Locationhttps://doi.org/10.1007/978-1-4939-7204-3_13
Isolation of newly transcribed RNA is an invaluable approach that can be used to study the dynamic life of RNA in cellulo. Traditional methods of whole-cell RNA extraction limit subsequent gene expression analyses to the steady-state levels of RNA abundance, which often masks changes in RNA synthesis and processing. This chapter describes a methodology with low cytotoxicity that permits the labeling and isolation of nascent pre-mRNA in cell culture. The resulting isolate is suitable for use in a series of downstream applications aimed at studying changes in RNA synthesis, processing, or stability.