Methods to determine mRNA half-life in Saccharomyces cerevisiae.
Much of our understanding of eukaryotic mRNA decay has come from studies in budding yeast, Saccharomyces cerevisiae. The facile nature of genetic and biochemical manipulations in yeast has allowed detailed investigations into the mRNA decay pathway and the identification of critical factors required for each step. Discussed herein are standard protocols for measuring mRNA half-lives in yeast. It should be noted, however, that variations of these assays are possible. In addition, a few a priori considerations are addressed.
Coller
Jeff
J
Center for RNA Molecular Biology, Case Western Reserve University, School of Medicine, Cleveland Ohio, USA.
D000483:Alleles; D000818:Animals; D015152:Blotting, Northern; D006207:Half-Life; D011401:Promoter Regions, Genetic / Q000235:genetics; D011760:Pyrrolidinones; D012319:RNA Polymerase II / Q000378:metabolism; D020871:RNA Stability / Q000235:genetics; D012331:RNA, Fungal / Q000032:analysis* / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D013696:Temperature
D011760:Pyrrolidinones; D012331:RNA, Fungal; C006361:acetopyrrothine; D012319:RNA Polymerase II