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High throughput genome methylated DNA enrichment method and indexes and index adapters used in same

A genomic and high-throughput technology, applied in the fields of MeDIP-seq library, tag technology, second-generation sequencing technology, and solexa sequencing technology, can solve the problem of time, labor and capital cost, feasibility discount, and complicated operation steps. Large-scale sample research and other issues, to achieve the effect of saving sample preparation time and reagent consumption, and low cost

Active Publication Date: 2014-05-14
BGI TECH SOLUTIONS
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Problems solved by technology

[0004] At present, the main defect of the existing MeDIP-seq technology is that the operation steps are cumbersome and it is difficult to study large-scale samples
The existing method is to carry out immune reaction on a single sample alone and then combine the enriched DNA with high-throughput sequencing research. This method makes the time, manpower and money spent on large sample size processing extremely huge, and is feasible. Big discount

Method used

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  • High throughput genome methylated DNA enrichment method and indexes and index adapters used in same
  • High throughput genome methylated DNA enrichment method and indexes and index adapters used in same
  • High throughput genome methylated DNA enrichment method and indexes and index adapters used in same

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Embodiment Construction

[0051] Embodiments of the present invention will be described in detail below in conjunction with examples, but those skilled in the art will understand that the following examples are only used to illustrate the present invention, and should not be considered as limiting the scope of the present invention.

[0052] Using this inventive method, we started with 6 samples of human peripheral blood genomic DNA (2 μg each) to construct a mixed library of 6 samples. TA clone was used to detect the quality of the library, and then a comparative analysis of high-throughput sequencing was carried out.

[0053] 1. Experimental part:

[0054] List of main experimental instruments

[0055]

[0056] List of related reagents

[0057]

[0058]

[0059] Table 1, the list of tags (DNA Index-N) and their corresponding tag adapters (composed of forward sequence DNA index-NF_adapter and reverse sequence DNA index-NR_adapter)

[0060]

[0061]

[0062]

[0063]

[0064] Ta...

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Abstract

Provided are DNA tags for constructing a MeDIP-seq library, a MeDIP-seq library and a method for preparing the same, detection method for the methylation of a plurality of DNA samples, and a kit for constructing the MeDIP-seq library. The DNA tags consist of the nucleotides shown in SEQ ID NO: (3N-2), in which N is any integer from 1 to 20.

Description

technical field [0001] The invention relates to the technical field of high-throughput genome methylation DNA enrichment. In addition, the present invention also relates to labeling technology and a method for constructing a label library, especially a MeDIP-seq library, in the same reaction system for multiple samples. The method of the present invention is particularly suitable for the second generation sequencing technology, especially the solexa sequencing technology. Background technique [0002] At present, genomic DNA methylation is one of the hottest directions in the field of epigenetics research, and it is gradually becoming an epigenetic marker for various diseases such as mammalian development and cancer. DNA methylation not only plays an important role in the modification of chromatin structure and genome stability, but also in eukaryotes, DNA methylation participates in various biological processes, such as embryonic development, genome imprinting, X chromosom...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C40B40/06C40B50/06C12Q1/68
CPCC40B70/00C40B50/06C12N15/1065C40B40/06
Inventor 孙继华王君文罗慧娟闫淑静章文蔚
Owner BGI TECH SOLUTIONS
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