Hi-C high-throughput sequencing library building method suitable for watermifoil

Abstract

The invention relates to a Hi-C high-throughput sequencing library building method suitable for watermifoil. The method includes the steps of firstly, performing formaldehyde crosslinking on a watermifoil sample to obtain a crosslinking material; secondly, performing cell lysis to release chromatin; thirdly, collecting the chromatin, and sequentially subjecting the chromatin to SDS deactivation endogenous enzyme and MboI endonuclease digestion breaking to obtain a material after digestion; fourthly, performing tail end filling to obtain tail-end-filled DNA, and performing intramolecular connection on the tail-end-filled DNA; fifthly, decrosslinking the intramolecular connection material and purifying DNA, and performing DNA breaking to form fragments beneficial to sequencing; seventhly, building a Hi-C library for high-throughput sequencing; eighthly, specifically capturing biotin marked DNA fragments, and performing amplification; ninthly, performing library quality inspection. The method has the advantages that the appropriate cell breaking method, formaldehyde crosslinking condition, endogenous enzyme deactivation condition and appropriate tool enzymes are designed according tothe special structure of watermifoil cells, the problem that the cell wall structure of the watermifoil affects a Hi-C experiment, Hi-C high-throughput sequencing library building of the watermifoil is achieved, and the application range of the Hi-C technology is expanded.

Description

technical field [0001] The present invention relates to the technical field of molecular biology, and more specifically relates to a Hi-C high-throughput sequencing library construction method suitable for Foxtail algae. Background technique [0002] High-throughput sequencing technology, also known as next-generation sequencing technology, is characterized by the ability to sequence hundreds of thousands to millions of DNA molecules in parallel at a time and the short read length. The second-generation high-throughput Illumina sequencing platform has the advantages of high sequencing throughput, high accuracy and low cost, and is widely used in many fields. [0003] Chromosome Conformation Capture (3C) technology is a technique for studying chromosome-protein interaction and chromosome conformation, which can provide detailed information on the association between distant genetic loci, which can be obtained from formaldehyde captured in the nucleus and can be deduced from ...

AI Technical Summary

Problems solved by technology

However, conventional Hi-C high-throughput sequencing library construction takes cell lines as the research object, and its chromatin is relatively simple. Although it can obtain better results, it limits its application range. For example, Foxtail algae is different from cell lines. The cell wall is rich in loose vacuolar tissue, and the cell nucleus cannot be released by simply lysing cells with plants; at the same time, experiments have found that poor endogenous enzyme inactivation will also affect the effect of subsequent enzyme digestion reactions, thereby affecting the validity of the library data

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