Blanshard Robert C, Chen Chongyi, Xie Xiaoliang Sunney, Hoffmann Eva R
Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Brighton, United Kingdom; Clinical Genomics Group, Illumina Inc., Fulbourn, United Kingdom.
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, United States.
Methods Cell Biol. 2018;144:441-457. doi: 10.1016/bs.mcb.2018.03.037. Epub 2018 Apr 24.
Genomic and chromosomal changes occur with a high rate in the germline and preimplantation embryos. To study such changes directly in the germline of mammals requires access to material as well as single cell genomics. Recent improvements in embryology and single-cell DNA amplification make it possible to study the genomic changes directly in human oocytes, sperm, and preimplantation embryos. This is particularly important for the study of chromosome segregation directly in human oocytes and preimplantation embryos. Here, we present a practical approach how to obtain high quality DNA sequences and genotypes from single cells, using manual handling of the material that makes it possible to detect genomic changes in meiosis and mitosis spanning the entire range from single nucleotide changes to whole chromosome aneuploidies.
基因组和染色体变化在种系和植入前胚胎中发生率很高。要直接在哺乳动物的种系中研究此类变化，需要获取材料以及进行单细胞基因组学研究。胚胎学和单细胞DNA扩增技术的最新进展使得直接在人类卵母细胞、精子和植入前胚胎中研究基因组变化成为可能。这对于直接在人类卵母细胞和植入前胚胎中研究染色体分离尤为重要。在此，我们提出一种实用方法，即如何通过手动处理材料从单细胞中获得高质量的DNA序列和基因型，从而能够检测从单核苷酸变化到整条染色体非整倍性的减数分裂和有丝分裂过程中的基因组变化。
