Biomedical Analysis Center, Army Medical University, Chongqing, China.
R&D Department, TCRCure Ltd., Chongqing, China.
J Mol Biol. 2019 Dec 6;431(24):5075-5085. doi: 10.1016/j.jmb.2019.08.002. Epub 2019 Sep 3.
High-throughput sequencing for transcriptome profiling is an increasingly accessible and important tool for biological research. However, accurate profiling of small cell populations remains challenging due to issues with gene detection sensitivity and experimental complexity. Here we describe a streamlined RNAseq protocol (EASY RNAseq) for sensitive transcriptome assessment starting from low amount of input materials. EASY RNAseq is technically robust enough for sequencing small pools of cells, recovering information on larger amounts of genes and with a more even expression distribution pattern than other commonly used methods. Application of EASY RNAseq to single-human embryos at the 8-cell stage led to detection of 70% of currently annotated protein-coding genes. This workflow may thus serve as a useful tool for sensitive interrogation of rare cell populations.
高通量测序进行转录组分析是生物学研究中一种越来越普及和重要的工具。然而，由于基因检测灵敏度和实验复杂性等问题，对小细胞群体的精确分析仍然具有挑战性。在这里，我们描述了一种简化的 RNAseq 方案（EASY RNAseq），用于从低量的输入材料开始进行敏感的转录组评估。EASY RNAseq 技术足够稳健，可以对小细胞池进行测序，恢复更多基因的信息，并且表达分布模式比其他常用方法更加均匀。将 EASY RNAseq 应用于 8 细胞期的单个人类胚胎，检测到了目前注释的 70%的蛋白编码基因。因此，该工作流程可以作为一种有用的工具，用于对稀有细胞群体进行敏感分析。
