High-resolution analysis of chromosome conformation in hyperthermophilic archaea.

High-resolution analysis of chromosome conformation in hyperthermophilic archaea.
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DOI:
10.1016/j.xpro.2021.100562
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发表时间:
2021-06-18
期刊:
影响因子:
--
通讯作者:
Bell SD
Bell SD
中科院分区:
其他
文献类型:
--
作者:
Takemata N;Bell SD

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染色体构象捕获(3C)技术正在成为研究古生代基因组组织的一种有前途的方法,古生代是染色体生物学中最不被理解的生命领域。在这里,我们描述了一种结合深度测序的3C技术,用于高温古生菌属Sulfolobus。该协议不使用与填充标记兼容的限制性内切酶,而是使用4个碱基的钝化切割器Alui来生成来自Sulfolobus物种的高分辨率(高达2kb)的联系人地图。有关本议定书的使用和执行的完整细节,请参阅。古硫柳的高分辨染色体构象捕获用于硫叶树DNA消化、连接和恢复过程的培养条件下一代测序的文库准备染色体构象捕获(3C)技术正在成为研究古生代基因组组织的一种有前途的方法,古生代是染色体生物学中了解最少的生命领域。在这里,我们描述了一种结合深度测序的3C技术,用于高温古生菌属Sulfolobus。该协议不使用与填充标记兼容的限制性内切酶,而是使用4个碱基的钝化切割器Alui来生成来自Sulfolobus物种的高分辨率(高达2kb)的联系人地图。
Chromosome conformation capture (3C) techniques are emerging as promising approaches to study genome organization in Archaea, the least understood domain of life in terms of chromosome biology. Here, we describe a 3C technique combined with deep sequencing for the hyperthermophilic archaeal genus Sulfolobus. Instead of using restriction enzymes compatible with fill-in labeling, this protocol uses the 4-bp blunt cutter AluI to generate high-resolution (up to 2 kb) contact maps from Sulfolobus species. For complete details on the use and execution of this protocol, please refer to. High-resolution chromosome conformation capture of Sulfolobus archaea Culture conditions for Sulfolobus DNA digestion, ligation, and recovery procedures Library preparation for next-generation sequencing Chromosome conformation capture (3C) techniques are emerging as promising approaches to study genome organization in Archaea, the least understood domain of life in terms of chromosome biology. Here, we describe a 3C technique combined with deep sequencing for the hyperthermophilic archaeal genus Sulfolobus. Instead of using restriction enzymes compatible with fill-in labeling, this protocol uses the 4-bp blunt cutter AluI to generate high-resolution (up to 2 kb) contact maps from Sulfolobus species.
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