Department of Biology, University of Florida, Gainesville, FL, United States.
Institute of Basic Medicine and Cancer (IBMC), Chinese Academy of Science, Hangzhou, China.
Methods Enzymol. 2023;683:153-170. doi: 10.1016/bs.mie.2022.08.039. Epub 2022 Nov 11.
Multi-omics has gained momentum over the past few years especially in plant single cell-type analysis as they aim to understand cellular molecular networks across different levels of genetic information flow. For multi-omics sample preparation, molecular extractions performed non-simultaneously create rooms for variation, inaccurate data, waste of limited samples, resources and labor. Here we optimized a protocol for 3-in-1 simultaneous extraction of RNA, metabolites, and proteins from the same single cell-type sample. We adapted a commercially available RNA kit with a few modifications to obtain high quality starting materials for sequencing and LC-MS/MS-based metabolomics and proteomics. RNAs are bound to the column, metabolites were extracted in a polar solvent and proteins are precipitated using acetone. This creates an all-in-one workflow using a standard RNA kit. Little training is required to carry out this protocol as it is simple and easy to use. It may be used with a wide range of plant species and different amounts of starting materials, including single cells.
多组学在过去几年中得到了迅猛发展，尤其是在植物单细胞类型分析中，因为它们旨在从不同层次的遗传信息流角度理解细胞分子网络。对于多组学样本制备，非同步进行的分子提取会产生变化的空间，导致数据不准确、有限样本、资源和劳动力的浪费。在这里，我们优化了一种从同一单细胞类型样本中同时提取 RNA、代谢物和蛋白质的 3-in-1 方法。我们对市售的 RNA 试剂盒进行了一些修改，以获得高质量的起始材料，用于测序和基于 LC-MS/MS 的代谢组学和蛋白质组学。RNA 与柱子结合，代谢物在极性溶剂中提取，蛋白质用丙酮沉淀。这使用标准 RNA 试剂盒创建了一个一体化工作流程。由于该方法简单易用，因此执行此方案所需的培训很少。它可以与多种植物物种和不同数量的起始材料（包括单细胞）一起使用。
