Department of Bioengineering, Stanford University, Stanford, CA 94305, USA.
Cell. 2012 Jul 20;150(2):402-12. doi: 10.1016/j.cell.2012.06.030.
Meiotic recombination and de novo mutation are the two main contributions toward gamete genome diversity, and many questions remain about how an individual human's genome is edited by these two processes. Here, we describe a high-throughput method for single-cell whole-genome analysis that was used to measure the genomic diversity in one individual's gamete genomes. A microfluidic system was used for highly parallel sample processing and to minimize nonspecific amplification. High-density genotyping results from 91 single cells were used to create a personal recombination map, which was consistent with population-wide data at low resolution but revealed significant differences from pedigree data at higher resolution. We used the data to test for meiotic drive and found evidence for gene conversion. High-throughput sequencing on 31 single cells was used to measure the frequency of large-scale genome instability, and deeper sequencing of eight single cells revealed de novo mutation rates with distinct characteristics.
减数分裂重组和新突变是配子基因组多样性的两个主要贡献因素，关于个体基因组如何被这两个过程编辑，仍有许多问题尚未解决。在这里，我们描述了一种用于单细胞全基因组分析的高通量方法，该方法用于测量一个个体配子基因组中的基因组多样性。微流控系统用于高度并行的样本处理，并最小化非特异性扩增。91 个单细胞的高密度基因分型结果用于创建个人重组图谱，该图谱与低分辨率的全人群数据一致，但在更高分辨率下与系谱数据显示出显著差异。我们使用这些数据来测试减数分裂驱动，并发现了基因转换的证据。对 31 个单细胞进行高通量测序以测量大规模基因组不稳定性的频率，对 8 个单细胞进行更深层次的测序揭示了具有独特特征的新突变率。
