IIGM Foundation - Italian Institute for Genomic Medicine, Torino, Italy.
Candiolo Cancer Institute, FPO-IRCCS, Torino, Italy.
Methods Mol Biol. 2022;2421:217-229. doi: 10.1007/978-1-0716-1944-5_15.
Over the past 7 years, single-cell sequencing has become very popular. For this reason, many laboratories of different biological disciplines that span from neurobiology to developmental biology from immunology to tumor biology have been approaching this technique. For someone new to this field that wants to investigate heterogeneity in what appears to be a single-cell population, the choice of the best protocol can be difficult, due to the high abundance of available protocols, instruments, and options. For this reason, here we describe the Smart-seq2 protocol for full-length mRNA sequencing of single cell. This protocol can be easily optimized in every molecular biology laboratory provided with standard laboratory equipment. The protocol is suitable for many different cell types, and the cost per cell is relatively small, allowing a good balance between costs and transcript coverage.
在过去的 7 年中，单细胞测序技术变得非常流行。出于这个原因，许多不同生物学领域的实验室，从神经生物学到发育生物学，从免疫学到肿瘤生物学，都在尝试这种技术。对于那些刚接触这个领域，想要研究似乎是单细胞群体中的异质性的人来说，由于可用的方案、仪器和选项数量众多，选择最佳方案可能会很困难。出于这个原因，在这里我们描述了 Smart-seq2 方案，用于单细胞全长 mRNA 测序。这个方案可以在配备标准实验室设备的每个分子生物学实验室中轻松优化。该方案适用于许多不同的细胞类型，并且每个细胞的成本相对较小，在成本和转录物覆盖之间实现了良好的平衡。
