Crutchfield Christopher A, Lu Wenyun, Melamud Eugene, Rabinowitz Joshua D
Lewis-Sigler Institute for Integrative Genomics, Department of Chemistry, Princeton University, Princeton, New Jersey, USA.
Methods Enzymol. 2010;470:393-426. doi: 10.1016/S0076-6879(10)70016-1. Epub 2010 Mar 1.
Driven by the advent of metabolomics, recent years have seen renewed interest in the investigation of yeast metabolism. Here we provide a practical guide to metabolomic analysis of yeast using liquid chromatography-mass spectrometry (LC-MS). We begin with background on LC-MS and its utility in studying yeast metabolism. We then describe key issues involved at each step of a typical yeast metabolomics experiment: in experimental design, cell culture, metabolite extraction, LC-MS, and data processing and analysis. Throughout, we highlight interdependencies between the steps that are relevant to developing an integrated workflow which effectively leverages LC-MS to reveal yeast biology.
在代谢组学出现的推动下,近年来人们对酵母代谢的研究重新产生了兴趣。在这里,我们提供了一份使用液相色谱 - 质谱联用(LC-MS)对酵母进行代谢组学分析的实用指南。我们首先介绍LC-MS的背景及其在研究酵母代谢中的作用。然后,我们描述了一个典型的酵母代谢组学实验每个步骤中涉及的关键问题:实验设计、细胞培养、代谢物提取、LC-MS以及数据处理和分析。在整个过程中,我们强调了与开发有效利用LC-MS揭示酵母生物学的综合工作流程相关的步骤之间的相互依存关系。