Chromosome conformation capture assay combined with biotin enrichment for hyperthermophilic archaea.

Chromosome organization in archaea has long been enigmatic due, in part, to the typically small cell size of archaea and the extremophilic nature of many of the model archaeal species studies, rendering live-cell imaging technically challenging. To circumvent these problems, we recently applied chromosome conformation capture combined with biotin enrichment and deep sequencing (Hi-C) to members of hyperthermophilic archaeal genus Sulfolobus. Our optimized Hi-C protocol described here permits delineation of how Sulfolobus species organize their chromosomes. For complete details on the use and execution of this protocol, please refer to. Growth of Sulfolobus species Cross-linking, digestion, and biotin end labeling DNA recovery, polishing, and biotin enrichment Library preparation and DNA sequencing Chromosome organization in archaea has long been enigmatic due, in part, to the typically small cell size of archaea and the extremophilic nature of many of the model archaeal species studies, rendering live-cell imaging technically challenging. To circumvent these problems, we recently applied chromosome conformation capture combined with biotin enrichment and deep sequencing (Hi-C) to members of hyperthermophilic archaeal genus Sulfolobus. Our optimized Hi-C protocol described here permits delineation of how Sulfolobus species organize their chromosomes.

古菌的染色体组织长期以来一直是个谜，部分原因是古菌通常细胞较小，而且许多作为研究模型的古菌物种具有极端嗜性，这使得活细胞成像在技术上具有挑战性。为了规避这些问题，我们最近将结合生物素富集和深度测序的染色体构象捕获技术（Hi - C）应用于超嗜热古菌属硫化叶菌的成员。我们在此描述的优化的Hi - C方案能够描绘硫化叶菌物种如何组织它们的染色体。 关于该方案的使用和实施的完整细节，请参考…… 硫化叶菌的生长 交联、酶切和生物素末端标记 DNA回收、修饰和生物素富集 文库制备和DNA测序 古菌的染色体组织长期以来一直是个谜，部分原因是古菌通常细胞较小，而且许多作为研究模型的古菌物种具有极端嗜性，这使得活细胞成像在技术上具有挑战性。为了规避这些问题，我们最近将结合生物素富集和深度测序的染色体构象捕获技术（Hi - C）应用于超嗜热古菌属硫化叶菌的成员。我们在此描述的优化的Hi - C方案能够描绘硫化叶菌物种如何组织它们的染色体。