Introducing Omni-C™ Technology
Dovetail™ Omni-C™ Technology incorporates a sequence-independent endonuclease approach to chromatin fragmentation in the proximity ligation protocol. This provides the following key advantages over traditional Hi-C that rely on restriction enzyme fragmentation:
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Genome wide coverage that is free of restriction enzyme density bias
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Shotgun-like coverage that captures SNPs, SVs and chromatin topology in a single NGS library
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Support for sample inputs down to 1,000 cells
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Compatible with hybrid capture approaches for targeted studies
By employing an endonuclease. Omni-C™ increases the genomic coverage of a proximity-ligation assay. therefore expanding the efficiency of each sequencing run by covering more of the genome and reducing biases imposed by RE site density As such. Omni-C™ generates libraries where more of the genome is included in analyses and thereby making the data more versatile and unbiased by RE sites.
Technology
Endonuclease-based molecular biology
The Omni-C™ process starts with endogenous chromatin being fixed in place (cross-linked) to create a stabilized nucleosome scaffold. A sequence independent endonuclease digests the cross-linked chromatin in situ. Upon release of the digested chromatin from the cell, free ends are ligated in a two step process that incorporates a biotinylated bridge between chromatin ends. The probability of two chromatin ends ligating together is dependent upon their proximity to each other within the scaffold. Following ligation and cross-link reversal, ligation products are enriched and sequenced using Illumina paired-end chemistry.

Process
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Omni-C™ Kit
The Omni-C™ Kit is currently validated for mammalian cells and tissues. The assay has a 2-day workflow from sample to sequencing-ready NGS library – sample prep and proximity ligation is completed on day 1 and library generation on day 2.
The assay incorporates three molecular biology-based quality control checks to enable identification of suboptimal libraries prior to sequencing. The 8-reaction Omni-C™ Kit provides enzymes and buffers necessary to complete day 1. The Dovetail ™ Library Module for Illumina or a paired-end sequencing kit for Illumina sequencing of your choosing is required for day 2. Omni-C™ data is compatible with a variety of open-source analysis and visualization tools.
- Prepare & Crosslink Sample (1-1.5 hrs) *optional stopping 🛑point
- In situ Nuclease Digestion (30 min)
- Quantify & Select Lysate (2 hrs) *optional stopping 🛑point
- Bind Chromatin to Chromatin Capture Beads (20 min)
- End Polishing (1 hr)
- Bridget Ligation (30 min)
- Intra-Aggregate Ligation (1 hr)
- Crosslink Reversal (1 hr)
- DNA Purification & Size Selection (30 min) *optional stopping 🛑point
- End Repair (1.25 hrs)
- Adapter Ligation (45 min)
- DNA Purification (30 min) *optional stopping 🛑point
- Ligation Capture (45 min)
- Index PCR (30 min)
- DNA Purification & Size Selection (30 min) *optional stopping 🛑point