DNA methylation detection method based on graphene oxide and restriction enzyme and kit thereof

A technology of restriction endonuclease and detection method, which is applied in biochemical equipment and methods, microbial determination/inspection, material excitation analysis, etc., to achieve the effect of simple operation and simple and fast detection method

Inactive Publication Date: 2014-01-15
SOUTHEAST UNIV
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Problems solved by technology

This DNA modification does not change the gene sequence, but it regulates gene expression

Method used

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  • DNA methylation detection method based on graphene oxide and restriction enzyme and kit thereof
  • DNA methylation detection method based on graphene oxide and restriction enzyme and kit thereof
  • DNA methylation detection method based on graphene oxide and restriction enzyme and kit thereof

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Embodiment 2

[0048] Example 2 The process of using a detection kit to detect DNA methylation

[0049] DNA Methylation Detection Kit:

[0050] 1) 25 μg / ml graphene oxide;

[0051] 2) restriction endonuclease HpaII;

[0052] 3) Probe DNA containing a CCGG sequence modified with a fluorescent group at one end;

[0053] 4) There is a segment of target gene DNA complementary to the probe DNA.

[0054] 1. Detection of DNA methylation by methyltransferase:

[0055] In 33mM tris-acetic acid (10mM Mg 2+ , 66mM Na + ) buffer system with a concentration of 1 μM, hybridize the DNA sequence of the methylated target gene caused by the methyltransferase with the probe DNA modified with a fluorescent group at one end containing the CCGG sequence; gradually add 25 μg / ml Graphene oxide, and then add restriction endonuclease HpaII, observe the degree of fluorescence recovery after double-stranded DNA interacts with graphene oxide.

[0056] 2. Detection of DNA methylation caused by chemical reagents: ...

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Abstract

The invention discloses a DNA methylation detection method. The DNA methylation detection method comprises the following steps: 1) selecting a DNA sequence; 2) hybridizing DNA double strands. The invention further discloses a DNA methylation detection kit. Through direct hybridization of the DNA strands and by virtue of interaction between the graphene oxide and the DNA, cytosine can be methylated in CpG island region through various methylation processes; through specific position recognizing and CpG position cutting functions of the restriction enzyme, an aim of detecting the methylated DNA is achieved; compared with the existing detection technology, the DNA methylation detection method has the characteristics of simpleness, fastness and convenience in operation; by the DNA methylation detection method, the effect of chemicals harmful to a human body on the DNA, namely DNA methylation, can be detected.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a DNA methylation detection method based on graphene oxide and restriction endonuclease and a kit thereof. Background technique [0002] DNA methylation is one of the earliest discovered gene epigenetic modifications, which may exist in all higher organisms. DNA methylation can turn off the activity of certain genes. The main forms of methylation are 5-methylcytosine, N6-methyladenine and 7-methylguanine. CCA / TGG and GATC are often methylated in prokaryotes, while methylation occurs only in cytosine in eukaryotes. Higher eukaryotic cells usually carry out methylation modification on the cytosine of the 5'-CpG-3' sequence on the DNA molecule, so that the cytosine at the 5' end of the CpG dinucleotide is converted into 5-methylcytosine. This DNA modification does not change the gene sequence, but it regulates gene expression. [0003] DNA methylation pla...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/44G01N21/64
CPCC12Q1/683C12Q2521/301
Inventor 卫伟熊艳翔刘松琴
Owner SOUTHEAST UNIV
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