Department of Data Science, Dana-Farber Cancer Institute, Boston, MA 02215, USA.
Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA.
STAR Protoc. 2021 Jul 17;2(3):100673. doi: 10.1016/j.xpro.2021.100673. eCollection 2021 Sep 17.
In many biological applications, the readout of somatic mutations in individual cells is essential. For example, it can be used to mark individual cancer cells or identify progenies of a stem cell. Here, we present a protocol to perform single-cell RNA-seq and single-cell amplicon-seq using 10X Chromium technology. Our protocol demonstrates how to (1) isolate CD34+ progenitor cells from human bone marrow aspirate, (2) prepare single-cell amplicon libraries, and (3) analyze the libraries to assign somatic mutations to individual cells. For complete details on the use and execution of this protocol, please refer to Van Egeren et al. (2021).
在许多生物应用中，对单个细胞中的体细胞突变进行读出是至关重要的。例如，它可用于标记单个癌细胞或鉴定干细胞的后代。在这里，我们展示了使用 10X Chromium 技术进行单细胞 RNA-seq 和单细胞扩增子测序的方案。我们的方案说明了如何：（1）从人骨髓抽吸物中分离 CD34+祖细胞，（2）制备单细胞扩增子文库，以及（3）分析文库，将体细胞突变分配给单个细胞。有关此方案的使用和执行的完整详细信息，请参阅 Van Egeren 等人（2021 年）。
