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Method for detecting DNMT1 based on self-assembled nucleic acid probe signal amplification method

A nucleic acid probe and self-assembly technology, applied in the field of biotechnology detection, can solve the problem that DNMT1DNMT1 is not included in cancer diagnosis

Pending Publication Date: 2021-04-09
ZHENGZHOU UNIV
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  • Abstract
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  • Application Information

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Problems solved by technology

However, neither DNMT1 activity nor DNMT1 levels have been incorporated into the indicators of cancer diagnosis

Method used

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  • Method for detecting DNMT1 based on self-assembled nucleic acid probe signal amplification method
  • Method for detecting DNMT1 based on self-assembled nucleic acid probe signal amplification method
  • Method for detecting DNMT1 based on self-assembled nucleic acid probe signal amplification method

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Embodiment Construction

[0033] The technical solutions of the present invention will be described in further detail below through specific implementation methods.

[0034] The instruments and equipment used in the examples of the present invention are shown in Table 1, the reagents and materials used are shown in Table 2, and the nucleic acids used are all purchased from Shanghai Sangon Bioengineering Co., Ltd., and the nucleic acid sequences are specifically shown in Table 3; the experimental water is Milli -Q water (resistivity greater than 18.2 MΩcm).

[0035] Table 3 The instruments and equipment used in the embodiment of the present invention

[0036] equipment name Model and manufacturer Fluorescence Spectrometer FS5, Edinburgh Instruments, UK Electric constant temperature water tank PK-8D, Shanghai Jinghong Experimental Equipment Co., Ltd. pH meter pH213, Italy HANNA company centrifuge Centrifuge 5418, eppendorf, Germany Oven Oscillator CHA-S, Cha...

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Abstract

The invention provides a method for detecting DNMT1 based on a self-assembled nucleic acid probe signal amplification method, which mainly utilizes an EDC / sulfo-NHS activation method to construct immunomagnetic beads, and McAbDNMT1 on the immunomagnetic beads can specifically capture DNMT1 in a sample; a double-antibody sandwich immunomagnetic bead and a biotinylated double-antibody sandwich immunomagnetic bead are sequentially formed; a signal amplification probe Biotin-poly FAM is constructed by using a nucleic acid self-assembly technology; the Biotin-poly FAM and the biotinylated double-antibody sandwich immunomagnetic beads can be coupled together through streptavidin to form a fluorescent compound; the method for detecting the DNMT1 level is established by utilizing the characteristic that the fluorescent compound can emit fluorescence and the linear relationship between the fluorescence intensity and the concentration of DNMT1, and the method has the characteristics of simplicity in operation, good accuracy, higher sensitivity, suitability for detecting the DNMT1 level in a biological sample and the like.

Description

technical field [0001] The invention relates to the field of biotechnology detection, in particular to a method for detecting DNMT1 based on a self-assembled nucleic acid probe signal amplification method. Background technique [0002] DNA methylation is an important part of epigenetics and plays an important role in cell proliferation, body aging, cancer and other life activities. DNA methyltransferase (DNMT1, DNMT3a, DNMT3b) is an important modification enzyme in the methylation process, and its main function is to transfer the methyl group of S-adenosylmethionine (SAM) to the corresponding DNA sequence. DNA methylation on cytosine or adenine. DNMTs are also involved in important biological activities, such as maintaining chromosome stability, embryonic development, and cell differentiation. Animal studies have shown that DNMT1 - / - Mice are stunted and die in the second trimester, DNMT3a - / - Mice died four weeks after birth, while DNMT3b - / - Mice died before birth. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6804G01N33/577G01N33/543G01N33/53
CPCC12Q1/6804G01N33/577G01N33/54326G01N33/5308C12Q2537/164C12Q2563/107C12Q2563/131
Inventor 于斐巩芳芳吴拥军刘利娥何磊良王艺琳万珍珍张咪咪宋露露杜梦思
Owner ZHENGZHOU UNIV
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