Hi-C is a comprehensive technique to capture the conformation of genomes. It is based on Chromosome Conformation Capture, where chromatin is crosslinked with formaldehyde, then digested and re-ligated so that only DNA fragments covalently linked together form ligation products. These products contain information about the genomic sequence and physical location in the 3D genome. Biotin-labeled nucleotides are incorporated at the ligation junction, allowing enrichment for chimeric DNA ligation junctions for deep sequencing. Hi-C is compatible with next-generation sequencing platforms, enabling unprecedented detection of chromatin interactions. This provides insights into chromatin biophysics and the implications of chromatin structure in nuclear functions. Hi-C offers a spatial context to genomic studies, enhancing understanding of chromatin's role in genome regulation in normal and disease conditions. Hi-C involves cell culture, crosslinking, lysis, digestion, biotin labeling, ligation, DNA purification, quality control, biotin removal, fragmentation, end repair, adapter ligation, and sequencing. The method ensures high complexity of the ligation product library, crucial for resolving chromatin interactions. Biotin labeling allows efficient purification of ligation products. DNA is fragmented and size-fractionated, followed by end repair and adapter ligation. The Hi-C library is then pulled down using streptavidin-coated beads, amplified, and sequenced. Hi-C data can be visualized and analyzed to reveal genome organization, chromatin compartmentalization, and chromosome territories. It provides a spatial context to biological inquiries, facilitating the discovery of gene regulation, nuclear partitioning, and chromatin dynamics. Hi-C offers a unique connectivity between genomic sequence and spatial conformation, making it ideal for studying genome organization and its implications for health and disease. Hi-C is compared to other methods like FISH and 5C, showing its advantages in genome-wide chromatin structure analysis. The method is supported by grants and acknowledges contributions from various researchers. Hi-C provides a powerful tool for studying nuclear organization and chromosome architecture.Hi-C is a comprehensive technique to capture the conformation of genomes. It is based on Chromosome Conformation Capture, where chromatin is crosslinked with formaldehyde, then digested and re-ligated so that only DNA fragments covalently linked together form ligation products. These products contain information about the genomic sequence and physical location in the 3D genome. Biotin-labeled nucleotides are incorporated at the ligation junction, allowing enrichment for chimeric DNA ligation junctions for deep sequencing. Hi-C is compatible with next-generation sequencing platforms, enabling unprecedented detection of chromatin interactions. This provides insights into chromatin biophysics and the implications of chromatin structure in nuclear functions. Hi-C offers a spatial context to genomic studies, enhancing understanding of chromatin's role in genome regulation in normal and disease conditions. Hi-C involves cell culture, crosslinking, lysis, digestion, biotin labeling, ligation, DNA purification, quality control, biotin removal, fragmentation, end repair, adapter ligation, and sequencing. The method ensures high complexity of the ligation product library, crucial for resolving chromatin interactions. Biotin labeling allows efficient purification of ligation products. DNA is fragmented and size-fractionated, followed by end repair and adapter ligation. The Hi-C library is then pulled down using streptavidin-coated beads, amplified, and sequenced. Hi-C data can be visualized and analyzed to reveal genome organization, chromatin compartmentalization, and chromosome territories. It provides a spatial context to biological inquiries, facilitating the discovery of gene regulation, nuclear partitioning, and chromatin dynamics. Hi-C offers a unique connectivity between genomic sequence and spatial conformation, making it ideal for studying genome organization and its implications for health and disease. Hi-C is compared to other methods like FISH and 5C, showing its advantages in genome-wide chromatin structure analysis. The method is supported by grants and acknowledges contributions from various researchers. Hi-C provides a powerful tool for studying nuclear organization and chromosome architecture.