Wellcome Trust-Medical Research Council, Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, Cambridge, UK.
Department of Haematology, University of Cambridge, Cambridge, UK.
Methods Mol Biol. 2021;2185:135-158. doi: 10.1007/978-1-0716-0810-4_9.
Single-cell RNA sequencing (scRNA-Seq) allows the complete and unbiased analysis of the transcriptional state of an individual cell. In the past 5 years, scRNA-Seq contributed to the progress of the hematology field, advancing our knowledge of both normal and malignant hematopoiesis. Different scRNA-Seq methods are available, all relying on the conversion of RNA to cDNA, followed by amplification of cDNA in order to obtain a sufficient amount of genetic material for sequencing. Currently available scRNA-Seq platforms can be broadly divided into two categories: droplet-based and plate-based. Each of these approaches has advantages and disadvantages that need to be considered when designing the experiment. Here, we describe detailed protocols of two of the most used methods for scRNA-Seq of hematopoietic cells: Smart-Seq2 (plate-based) and 10× Genomics (droplet-based).
单细胞 RNA 测序（scRNA-Seq）允许对单个细胞的转录状态进行完整和无偏的分析。在过去的 5 年中，scRNA-Seq 推动了血液学领域的进展，增进了我们对正常和恶性造血的认识。有不同的 scRNA-Seq 方法，都依赖于 RNA 向 cDNA 的转化，然后对 cDNA 进行扩增，以获得足够用于测序的遗传物质。目前可用的 scRNA-Seq 平台可大致分为两类：基于液滴的和基于平板的。在设计实验时，需要考虑到这些方法中的每一种都有其优点和缺点。在这里，我们描述了两种最常用于造血细胞 scRNA-Seq 的方法的详细方案：Smart-Seq2（基于平板）和 10× Genomics（基于液滴）。
