Small RNAs participate in several cellular processes, including splicing, RNA modification, mRNA degradation, and translational arrest. Traditional methods for sequencing small RNAs require a large amount of cell material, limiting the possibilities for single-cell analyses. We describe Small-seq, a ligation-based method that enables the capture, sequencing, and molecular counting of small RNAs from individual mammalian cells. Here, we provide a detailed protocol for this approach that relies on standard reagents and instruments. The standard protocol captures a complex set of small RNAs, including microRNAs (miRNAs), fragments of tRNAs and small nucleolar RNAs (snoRNAs); however, miRNAs can be enriched through the addition of a size-selection step. Ready-to-sequence libraries can be generated in 2-3 d, starting from cell collection, with additional days needed to computationally map the sequence reads and calculate molecular counts.

中文翻译：

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用于单细胞小RNA测序的Small-seq。

小RNA参与多个细胞过程，包括剪接，RNA修饰，mRNA降解和翻译停滞。用于对小RNA进行测序的传统方法需要大量的细胞材料，从而限制了单细胞分析的可能性。我们描述了Small-seq，这是一种基于连接的方法，能够从单个哺乳动物细胞中捕获，测序和对小RNA进行分子计数。在这里，我们为依赖标准试剂和仪器的这种方法提供了详细的协议。标准协议捕获了一组复杂的小RNA，包括microRNA（miRNA），tRNA片段和小核仁RNA（snoRNA）。但是，可以通过增加大小选择步骤来丰富miRNA。从细胞收集开始，可以在2-3 d内生成准备测序的文库，