Exploring DNA quality of single cells for genome analysis with simultaneous whole-genome amplification
Exploring DNA quality of single cells for genome analysis with simultaneous whole-genome amplification
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DOI：
10.1038/s41598-018-25895-7
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发表时间：
2018-05-10
影响因子：
4.6
通讯作者：
Korfhage, Christian
中科院分区：
文献类型：
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作者：
Baeumer, Christiane;Fisch, Evelyn;Korfhage, Christian
单细胞基因组分析方法是定义单细胞特征并识别它们之间差异的强大工具。由于单个细胞的DNA量非常有限，细胞DNA通常需要通过全基因组扩增的方式进行扩增，然后才能进行进一步分析。单个核仅包含两个单倍体基因组。因此，任何阻止扩增的 DNA 损伤都会导致受损 DNA 位点的丢失并引发扩增偏差。因此，迫切需要对单细胞DNA质量进行评估。迄今为止，还没有一种简单的方法可以确定单细胞 DNA 的质量，同时仍保留整个细胞 DNA 进行扩增和下游分析。在这里，我们描述了一种通过使用竞争性掺入 DNA 模板进行全基因组扩增并同时对单细胞 DNA 进行质量控制的方法。
Single cell genome analysis methods are powerful tools to define features of single cells and to identify differences between them. Since the DNA amount of a single cell is very limited, cellular DNA usually needs to be amplified by whole-genome amplification before being subjected to further analysis. A single nucleus only contains two haploid genomes. Thus, any DNA damage that prevents amplification results in loss of damaged DNA sites and induces an amplification bias. Therefore, the assessment of single cell DNA quality is urgently required. As of today, there is no simple method to determine the quality of a single cell DNA in a manner that will still retain the entire cellular DNA for amplification and downstream analysis. Here, we describe a method for whole-genome amplification with simultaneous quality control of single cell DNA by using a competitive spike-in DNA template.