van den Bos Hilda, Bakker Bjorn, Taudt Aaron, Guryev Victor, Colomé-Tatché Maria, Lansdorp Peter M, Foijer Floris, Spierings Diana C J
European Research Institute for the Biology of Ageing (ERIBA), University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
Institute for Computational Biology, Helmholtz Zentrum München, Neuherberg, Germany.
Methods Mol Biol. 2019;1896:159-190. doi: 10.1007/978-1-4939-8931-7_15.
High-throughput next generation sequencing karyotyping has emerged as a powerful tool for the detection of genomic heterogeneity in normal tissues and cancers. Here we describe a single-cell whole genome sequencing (scWGS) platform to assess whole-chromosome aneuploidy, structural aneuploidies involving only chromosome fragments and more local small copy number alterations in individual cells. We provide a detailed protocol for the isolation, library preparation, low coverage sequencing and data analysis of single cells. Since our approach does not involve a whole-genome preamplification step, our method allows for acquisition of reliable high-resolution single-cell copy number profiles. Moreover, the protocol allows multiplexing of 384 single-cell libraries in one sequencing run, thereby significantly reducing sequencing costs and can be completed in 3-4 days starting from single cell isolation to analysis of sequencing data.
高通量下一代测序核型分析已成为检测正常组织和癌症中基因组异质性的有力工具。在此，我们描述了一种单细胞全基因组测序（scWGS）平台，用于评估单个细胞中的全染色体非整倍性、仅涉及染色体片段的结构非整倍性以及更多局部小拷贝数改变。我们提供了单个细胞分离、文库制备、低覆盖度测序和数据分析的详细方案。由于我们的方法不涉及全基因组预扩增步骤，我们的方法能够获取可靠的高分辨率单细胞拷贝数图谱。此外，该方案允许在一次测序运行中对384个单细胞文库进行多重分析，从而显著降低测序成本，并且从单细胞分离到测序数据分析可在3 - 4天内完成。
