Deng Wulan, Blobel Gerd A
Transcription Imaging Consortium, Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, VA, 20147, USA.
Division of Hematology, The Children's Hospital of Philadelphia, Philadelphia, PA, 19104, USA.
Methods Mol Biol. 2017;1468:51-62. doi: 10.1007/978-1-4939-4035-6_6.
Chromosome conformation capture (3C) technology and its derivatives are currently the primary methodologies measuring contacts among genomic elements. In fact, the lion share of what is currently known about chromosome folding is based on 3C-related approaches. For example, distal enhancers are commonly in physically proximity with their target genes, forming chromatin loops. Additional layers of chromatin organization have been described using 3C-based techniques, including topological domains (TADs) and sub-TADs. Finally, inter-chromosomal interactions have been reported although they are much less frequent. 3C is becoming increasingly widespread in its use for understanding genome organization. Here we provide a protocol for quantitative 3C using real-time PCR analysis, along with essential quality controls and normalization methods.
染色体构象捕获（3C）技术及其衍生技术是目前测量基因组元件之间相互作用的主要方法。事实上，目前已知的关于染色体折叠的大部分信息都基于与3C相关的方法。例如，远端增强子通常与其靶基因在物理上相邻，形成染色质环。使用基于3C的技术已经描述了染色质组织的其他层次，包括拓扑结构域（TADs）和亚TADs。最后，尽管染色体间相互作用的频率要低得多，但也有相关报道。3C在用于理解基因组组织方面的应用越来越广泛。在此，我们提供了一种使用实时PCR分析进行定量3C的方案，以及基本的质量控制和标准化方法。
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