Leman Adam R, Bristow Sara L, Haase Steven B
Department of Biology, Duke University, 90338, Science Drive, Durham, NC, 27708, USA.
Methods Mol Biol. 2014;1170:295-312. doi: 10.1007/978-1-4939-0888-2_14.
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.
在芽殖酵母中测定全局细胞周期调控转录涉及从同步群体中提取RNA以及对检测到的转录水平进行适当标准化。在这里，我们描述了通过离心淘析使酿酒酵母细胞群体同步化，随后分离RNA用于微阵列分析的方法。此外，我们概述了直接比较实验中各个时间点的RNA丰度以及比较独立实验所需的计算方法。这些方法共同描述了在细胞周期中观察RNA丰度所需的完整工作流程。
