title: Analyzing transcription dynamics during the budding yeast cell cycle.
abstract: Assaying global cell cycle-regulated transcription in budding yeast involves extracting RNA from a synchronous population and proper normalization of detected transcript levels. Here, we describe synchronization of Saccharomyces cerevisiae cell populations by centrifugal elutriation, followed by the isolation of RNA for microarray analysis. Further, we outline the computational methods required to directly compare RNA abundance from individual time points within an experiment and to compare independent experiments. Together, these methods describe the complete workflow necessary to observe RNA abundance during the cell cycle.
authors.affiliation: Department of Biology, Duke University, 90338, Science Drive, Durham, NC, 27708, USA.
authors.forename: Adam R
authors.initials: AR
authors.lastname: Leman
authors.forename: Sara L
authors.initials: SL
authors.lastname: Bristow
authors.forename: Steven B
authors.initials: SB
authors.lastname: Haase
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods; D002453:Cell Cycle; D002469:Cell Separation / Q000379:methods; D002498:Centrifugation / Q000379:methods; D004586:Electrophoresis / Q000379:methods; D015966:Gene Expression Regulation, Fungal; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D012331:RNA, Fungal / Q000032:analysis / Q000235:genetics* / Q000302:isolation & purification; D004718:Saccharomycetales / Q000166:cytology* / Q000235:genetics* / Q000254:growth & development; D015533:Transcriptional Activation
chemical_list: D012331:RNA, Fungal