Identification of changes in gene expression by quantitation of mRNA levels.
Sequence elements within mRNA-untranslated regions and their binding partners are key controllers of mRNA stability. Changes in mRNA stability can often be detected by changes in steady-state mRNA abundance, or a more careful analysis of mRNA half-lives can be performed following transcriptional repression. This chapter presents methods to isolate RNA from both yeast and mammalian cells for either steady-state or half-life analyses. In addition, two reliable methods to quantitate mRNA levels, northern blot analysis and real-time PCR, are outlined and compared.
Olivas
Wendy M
WM
Department of Biology, University of Missouri - St. Louis, St. Louis, MO, USA.
D000818:Animals; D015152:Blotting, Northern / Q000379:methods; D004587:Electrophoresis, Agar Gel; D015870:Gene Expression; D006207:Half-Life; D006801:Humans; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods; D020871:RNA Stability; D012331:RNA, Fungal / Q000032:analysis / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism
D015345:Oligonucleotide Probes; D012331:RNA, Fungal; D012333:RNA, Messenger