Protocol to analyze steps of ribosome biogenesis in nuclear and cytosolic fractions of yeast cells
Abstract
Cell fractionation enables the study of compartment-specific RNA processing events in organisms with complex cell walls. Here, we present a protocol for isolating nuclear and cytosolic fractions from Saccharomyces cerevisiae . We describe spheroplast formation using zymolyase, followed by subcellular fractionation via a sucrose density gradient. Finally, we detail the extraction of high-quality RNA from these fractions. This approach overcomes cell wall barriers to allow the analysis of ribosome biogenesis. For complete details on the use and execution of this protocol, please refer to Zadali et al . 1
Fields of Study & Subjects
RNA and protein synthesis mechanisms
Nuclear Structure and Function
RNA Research and Splicing
Publication details
- Journal / source
- STAR Protocols
- Volume
- 7
- Issue
- 1
- Pages
- 104429–104429
- Publication date
- 1 March 2026
- Language
- EN