Time-of-flight mass spectrometry multi-target DNA methylation site quantitative detection method

A technology of time-of-flight mass spectrometry and quantitative detection methods, which is applied to biochemical equipment and methods, and microbial measurement/inspection. The effect of cost reduction and accurate results

Inactive Publication Date: 2021-04-30
博淼生物科技(北京)有限公司
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

The disadvantage of this detection method is that it is impossible to detect CpG sites that are not in the same range in the same reaction well at the same time, and because of technical limitations, it cannot be guaranteed that the CpG sites in the amplified fragments will be detected, or they will be detected separately. Test results

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  • Time-of-flight mass spectrometry multi-target DNA methylation site quantitative detection method
  • Time-of-flight mass spectrometry multi-target DNA methylation site quantitative detection method
  • Time-of-flight mass spectrometry multi-target DNA methylation site quantitative detection method

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Experimental program
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Embodiment Construction

[0045] The experimental procedure is attached figure 1 .

[0046] For convenience, the reagents required for the experiment are summarized as follows: EZ DNA Methylation Gold kit (ZYMO), ddH 2 O, PCR Buffer, MgCl 2, dNTPs, Taq Polymerase, SAP Buffer, Shrimp Alkaline Phosphatase Enzyme, iPLEX Buffer Plus, iPLEX Termination Mix, iPLEX Enzyme.

[0047] (1) The detection method of the present invention has a wide range of sample types, such as blood, tissue, cells, etc., and can efficiently extract and prepare genomic DNA for quantitative methylation detection.

[0048]

[0049] (2) CT treatment of DNA:

[0050] I. Reaction system

[0051] Reagent Volume for CT Conversion Reagent ddH2O 9mL M-Dilution Buffer 3mL M-Dissolving Buffer 500μL

[0052] II. Reaction procedure

[0053] temperature time number of cycles 98℃ 10min 1 64℃ 30min 5 4℃ ∞ 1

[0054] (3) Amplify the target fragment:

[0055] I, re...

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Abstract

The invention belongs to the technical field of biology, particularly discloses a time-of-flight mass spectrometry multi-target DNA methylation site quantitative detection method, and provides a high-throughput detection method of DNA methylation sites, which can design a specific probe for multiple methylation sites to be detected, and in the same reaction system, a quantitative methylation degree detection result is obtained. The method has the characteristics that the operation is simple, the methylation sites in different regions can be simultaneously detected, the corresponding experiment cost is reduced, and the like.

Description

technical field [0001] The invention belongs to the field of biological technology, and specifically discloses a time-of-flight mass spectrometry multiple targeting DNA methylation site quantitative detection method. [0002] technical background [0003] As an important chemical modification of DNA, DNA methylation can affect the genetic expression of DNA without changing the DNA sequence. In vertebrates, the vast majority of DNA methylation is modified to 5-methylcytosine (5mc). In the human genome, about 70% to 80% of the CpG dinucleotides are involved in the methylation of this modified DNA. Kylation can be involved in many biological processes, including genome imprinting, transposable element silencing, stem cell differentiation, embryonic development, etc. As an important epigenetic phenomenon, DNA methylation is one of the important research contents of epigenetics. [0004] DNA methylation detection methods are developing rapidly. High-throughput sequencing technol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6872C12Q1/6851
CPCC12Q1/6851C12Q1/6872C12Q2600/154
Inventor 张方舟廉欢欢张起起秦胜红王新新梁海泳
Owner 博淼生物科技(北京)有限公司
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