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DOI: 10.3791/56286-v
Please note that some of the translations on this page are AI generated. Click here for the English version.
This article describes a technique that combines flow cytometry and high throughput sequencing to identify late replicating regions of the genome. This method provides insights into the dynamics of genomic replication and helps address key questions in the DNA replication field.
我们描述了一种将流式细胞术和高通量测序相结合的技术来识别基因组的晚期复制区域。
该程序对根据细胞周期阶段进行流式分选的细胞的 DNA 进行深度测序的总体目标是识别在细胞周期中复制极晚的基因组区域。这种方法可以帮助回答 DNA 复制领域的关键问题,例如基因组的哪些区域难以完成 DNA 复制,因此特别容易受到基因组重排的影响。该技术的主要优点是,尽管从实验的角度来看非常简单,但它为基因组复制的动力学提供了令人惊讶的丰富视图。
首先,用感兴趣的酵母细胞接种 15 毫升含有 8 毫升 YEPD 的试管。合成或富介质是可以接受的。将细胞培养过夜至少 12 小时,以便在其实际对数期分布期间收集它们。
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