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Method for constructing DNA library and application of method

A technology of DNA library and three-dimensional structure, which is applied in the construction of DNA library and its application field, can solve the problems affecting the proportion of effective data, the loss of effective fragments, and the quality of the library, so as to achieve a high proportion of effective data, appropriate length of fragments, and time to build a library The effect of shortening

Pending Publication Date: 2019-12-24
安诺优达生命科学研究院 +4
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Problems solved by technology

[0002] There are many steps in the existing next-generation sequencing library construction technology, especially the multi-step operations from the final end repair to PCR are likely to cause the loss of effective fragments
It is more significant in the three-dimensional genome Hi-C library construction. The biotin-labeled chimeric DNA is relatively small as a library construction template, so the loss of effective fragments after fishing will directly affect the quality of the final library.
In addition, the fragment screening step is set to make the library adapt to the sequencing-by-synthesis principle of illumina, but too long fragments will lead to poor quality of sequencing data
At the same time, a considerable part of the available library fragments will be screened out due to fragment length issues, especially under the conditions of nanogram-level templates, screening will lead to a reduction in the number of effective libraries, directly affecting the effective data ratio
[0003] Therefore, existing library construction methods need to be improved

Method used

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  • Method for constructing DNA library and application of method
  • Method for constructing DNA library and application of method
  • Method for constructing DNA library and application of method

Examples

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Embodiment 1

[0064] Utilize the method of the embodiment of the present invention, take mouse cell as sample, construct DNA library, and carry out sequencing and quality control, specifically as follows:

[0065] 1. Experimental method

[0066] 1. Preparation of experimental materials

[0067] The mouse 3T3-NIH cell line was used as the experimental material. The frozen 3T3-NIH cells were quickly thawed in a 37°C water bath, and then transferred to 9ml cell culture medium in a biological safety cabinet and mixed evenly. The cell culture medium included 15% fetal bovine serum, 84% DMEM culture medium and 1% Streptomycin antibody (both V / V). Centrifuge at 1000rpm at 23°C for 10min, discard the supernatant, add 5ml of cell culture medium, resuspend the cells and transfer to a cell culture flask, place in a cell culture incubator at 37°C, 5% CO 2 Static cultivation.

[0068] When the cells grow to 80% full (about 1-2 days), take out the culture bottle, pour out the culture medium in a biol...

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Abstract

The invention discloses a method for constructing a DNA library and application of the method. The method for constructing the DNA library comprises the steps: providing DNA of a chimeric marker, wherein the DNA of the chimeric marker has three-dimensional structure information; carrying out transposition treatment on the DNA of the chimeric marker to obtain a transposition product; carrying out capture treatment on the transposition product to obtain captured DNA; and carrying out amplification treatment on the captured DNA so as to obtain the DNA library. The library construction method is simple in steps, short in consumed time, particularly suitable for library construction of trace DNA samples, high in effective data proportion of sequencing and low in noise single-end suspension value.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular, to a method for constructing a DNA library and its application, more specifically, to a method for constructing a DNA library, a method for obtaining chromatin interaction information in individual cells, and a method for obtaining A method for information, a method for three-dimensional genome research, a method for prenatal diagnosis or cancer screening, a kit, and the use of the kit in three-dimensional genome assembly library or prenatal diagnosis or cancer screening. Background technique [0002] There are many steps in the existing next-generation sequencing library construction technology, especially the multi-step operations from the final end repair to PCR are likely to cause the loss of effective fragments. It is more significant in the three-dimensional genomic Hi-C library construction. The biotin-labeled chimeric DNA is relatively small as a library construction te...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806C12N15/10C40B50/06
CPCC12N15/1093C12Q1/6806C40B50/06C12Q2525/191C12Q2563/131C12Q2531/113
Inventor 陈晓丹徐护朝潘伟业李志民李大为玄兆伶王海良王娟
Owner 安诺优达生命科学研究院
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