Sequencing the genomes of individual cells enables the direct determination of genetic heterogeneity amongst cells within a population. We have developed an injection-moulded valveless microfluidic device in which single cells from colorectal cancer derived cell lines (LS174T, LS180 and RKO) and fresh colorectal tumors have been individually trapped, their genomes extracted and prepared for sequencing using multiple displacement amplification (MDA). Ninety nine percent of the DNA sequences obtained mapped to a reference human genome, indicating that there was effectively no contamination of these samples from non-human sources. In addition, most of the reads are correctly paired, with a low percentage of singletons (0.17 ± 0.06%) and we obtain genome coverages approaching 90%. To achieve this high quality, our device design and process shows that amplification can be conducted in microliter volumes as long as the lysis is in sub-nanoliter volumes. Our data thus demonstrates that high quality whole genome sequencing of single cells can be achieved using a relatively simple, inexpensive and scalable device. Detection of genetic heterogeneity at the single cell level, as we have demonstrated for freshly obtained single cancer cells, could soon become available as a clinical tool to precisely match treatment with the properties of a patient's own tumor.

中文翻译：

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从无阀微流控设备中分离的单个癌细胞提取的人类基因组测序†

对单个细胞的基因组测序可以直接确定群体中细胞之间的遗传异质性。我们已经开发了一种无瓣膜注射微流控设备，其中来自结直肠癌的细胞系（LS174T，LS180和RKO）的单个细胞和新鲜的结直肠肿瘤已被单独捕获，提取了它们的基因组，并准备使用多重置换扩增（MDA）进行测序。所获得的DNA序列的百分之九十九映射到参考人类基因组，这表明这些样品实际上没有受到来自非人类来源的污染。此外，大多数读段均已正确配对，单例百分率较低（0.17±0.06％），而我们获得的基因组覆盖率接近90％。为了达到如此高的质量，我们的设备设计和过程表明，只要裂解液的体积在亚纳升以下，就可以在微升体积中进行扩增。因此，我们的数据表明，使用相对简单，便宜且可扩展的设备可以实现单细胞的高质量全基因组测序。正如我们已经证明的那样，对于新鲜获得的单个癌细胞，在单细胞水平上检测遗传异质性可能很快将成为一种临床工具，以使治疗与患者自身肿瘤的特性精确匹配。