title: Quantitative Northern Blot Analysis of Mammalian rRNA Processing.
abstract: Assembly of eukaryotic ribosomes is an elaborate biosynthetic process that begins in the nucleolus and requires hundreds of cellular factors. Analysis of rRNA processing has been instrumental for studying the mechanisms of ribosome biogenesis and effects of stress conditions on the molecular milieu of the nucleolus. Here, we describe the quantitative analysis of the steady-state levels of rRNA precursors, applicable to studies in mammalian cells and other organisms. We include protocols for gel electrophoresis and northern blotting of rRNA precursors using procedures optimized for the large size of these RNAs. We also describe the ratio analysis of multiple precursors, a technique that facilitates the accurate assessment of changes in the efficiency of individual pre-rRNA processing steps.
authors: Department of Cell Biology, Rowan University School of Osteopathic Medicine, 2 Medical Center Drive, Stratford, NJ, 08084, USA.|Blood Cell Development and Function Program, Fox Chase Cancer Center, Philadelphia, PA, USA.
authors: Minshi
authors: M
authors: Wang
authors: Department of Cell Biology, Rowan University School of Osteopathic Medicine, 2 Medical Center Drive, Stratford, NJ, 08084, USA. pestovdg@rowan.edu.
authors: Dimitri G
authors: DG
authors: Pestov
mesh_terms: D000818:Animals; D015152:Blotting, Northern / Q000379:methods; D006801:Humans; D012322:RNA Precursors / Q000235:genetics; D012323:RNA Processing, Post-Transcriptional; D012335:RNA, Ribosomal / Q000235:genetics*; D014158:Transcription, Genetic
chemical_list: D012322:RNA Precursors; D012335:RNA, Ribosomal
keywords: Hybridization; Mammalian cells; Northern blot; Pre-rRNA; RNA electrophoresis; Ribosome biogenesis