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A method for constructing hi-c libraries with high available data rate

A library and hi-c technology, applied in chemical libraries, biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problem of low data rate of Hi-C library and achieve the effect of increasing the available data rate

Active Publication Date: 2019-11-29
ZHEJIANG ANNOROAD BIO TECH CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, the Hi-C library constructed by the method reported in this literature has the problem of low available data rate

Method used

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  • A method for constructing hi-c libraries with high available data rate
  • A method for constructing hi-c libraries with high available data rate
  • A method for constructing hi-c libraries with high available data rate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Human leukocyte Hi-c library construction

[0074] 1. Formaldehyde crosslinking

[0075] 1. Formaldehyde crosslinking

[0076] 1.1) The cell type is human leukocytes, the number of cells is 5×10 6 cells. The cultured cell supernatant was discarded, and 22.5 mL of fresh serum-free culture solution was added to each bottle.

[0077] 1.2) Add 1.25mL of 37% formaldehyde to make the final concentration 2% by weight, shake quickly after adding formaldehyde.

[0078] 1.3) Accurately incubate at room temperature for 10 minutes. Shake the culture gently every 2 min.

[0079] 1.4) Add 2.5mL of 2.5M glycine to terminate cross-linking and mix well.

[0080] 1.5) Incubate at room temperature for 10 minutes, and then place on ice for 15 minutes to completely terminate the reaction.

[0081] 1.6) Collect the cells and transfer them into tubes.

[0082] 1.7) Collect cross-linked cells by centrifugation at 800×g for 10 min.

[0083] 1.8) Remove the supernatant.

[0084] 1.9) T...

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Abstract

The present invention provides a method for constructing a high available data rate Hi-C library. The method comprises: treating cells by using more than 1 wt% of a formaldehyde solution to obtain cells having immobilized chromatin; and washing a streptavidin-based solid carrier having an immobilized biotin-labeled DNA fragment by using a 0.01-1 M NaOH solution. With the method of the present invention, by cleverly improving on the basis of the traditional Hi-C library construction method, the available data rate of the constructed Hi-C library can be significantly improved.

Description

technical field [0001] The invention relates to a method for constructing a library capable of capturing the three-dimensional conformation of chromatin within the scope of the whole genome, and belongs to the technical field of gene sequencing. Background technique [0002] DNA is the carrier of cell genetic information, exists in each cell in the form of chromatin in organisms, and controls the process of the entire life activity. At present, the vast majority of research on DNA information is carried out by studying the sequence of bases in DNA molecules (one-dimensional information of DNA), and exploring the laws of life activities by analyzing base arrangement information. [0003] The nucleus in the real state is a narrow three-dimensional space. DNA with a linear molecular structure will be located in the nucleus in a complex coiled manner. The original one-dimensional DNA sequence is endowed with a three-dimensional spatial conformation, which leads to a large number...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10C12Q1/6869C40B50/06
Inventor 张介中李珊珊赵红梅苑赞玄兆伶李大为梁峻彬陈重建
Owner ZHEJIANG ANNOROAD BIO TECH CO LTD