Efficient whole-genome chromosome conformation capture technology (eHi-C)

A chromatin and ehi-c technology, which is applied in the direction of recombinant DNA technology, combinatorial chemistry, microbial measurement/testing, etc., can solve the problems of increased cost of cell culture, increased experimental error, and low resolution of interaction, etc. Effects of required amount of cells, saving reagents, and simplifying steps
CN106566828AActive Publication Date: 2017-04-19AGRI GENOMICS INST CHINESE ACADEMY OF AGRI SCI
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
AGRI GENOMICS INST CHINESE ACADEMY OF AGRI SCI
Publication Date
2017-04-19

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Abstract

The invention discloses an efficient whole-genome chromosome conformation capture technology (eHi-C). The invention provides an eHi-C sequencing technology for cells. The eHi-C sequencing technology comprises the following steps: 1) subjecting the cells to disintegration so as to obtain chromatin; 2) subjecting the chromatin to enzyme digestion, DNA end labeling, cyclization connection of a blunt end and DNA purification so as to obtain purified circular DNA; 3) introducing a circular co-precipitation DNA molecule used as internal reference; 4) carrying out ultrasonic breaking; 5) capturing all the labeled DNA fragments with immunomagnetic beads; and 6) preparing eHi-C sequencing library from the labeled DNA fragments. The technology provided by invention is low in the amount of needed cells, can easily acquire an experiment material, reduces time in acquisition of the material, greatly lowers cell culture cost and cost for reagent consumables and decreases test errors.
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Description

technical field

[0001] The invention relates to the technical field of genome sequencing, in particular to eHi-C, an efficient genome-wide chromatin conformation technology. Background technique

[0002] Three-dimensional (3D) genomics is a discipline that studies the three-dimensional spatial structure and function of the genome. Specifically, it refers to the study of genome sequence, gene structure and its regulatory elements, the three-dimensional spatial structure of genome sequence in the nucleus, and its function in biological processes such as gene transcription, replication, repair, and regulation. The related sequencing technologies of 3D genome mainly include Hi-C (High-throughput / resolution chromosome conformationcapture) developed by Job Dekker and ChIA-PET (Chromatin Interaction Analysis with Paired-End Tag Sequencing) technology developed by Professor Ruan Yijun (Li Guoliang, Ruan Yijun, 2014, Science Bulletin).

[0003] Hi-C technology is based on high-thro...

Claims

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