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Primer and kit for detecting methylation of MGMT (O<6>-methylguanine-DNA methyltransferase) genes and use method of primer

A methylation and kit technology, applied in the field of molecular biology, can solve the problems of low sensitivity and high detection cost of direct sequencing method, achieve high accuracy, high sensitivity, and reduce work intensity

Inactive Publication Date: 2014-02-12
北京海思特医学检验实验室有限公司
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Problems solved by technology

The sensitivity of the direct sequencing method is low, requiring the mutation ratio of the tested material to be greater than 20%, and the detection cost is also high

Method used

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  • Primer and kit for detecting methylation of MGMT (O&lt;6&gt;-methylguanine-DNA methyltransferase) genes and use method of primer
  • Primer and kit for detecting methylation of MGMT (O&lt;6&gt;-methylguanine-DNA methyltransferase) genes and use method of primer
  • Primer and kit for detecting methylation of MGMT (O&lt;6&gt;-methylguanine-DNA methyltransferase) genes and use method of primer

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Embodiment

[0023] Kit for detecting MGMT gene methylation and use thereof

[0024] 1. The kit for MGMT gene methylation contains:

[0025] (1) primers, the primer sequences are shown in SEQ ID NO: 1 to SEQ ID NO: 6, and the concentration of the above primers is 10 pmol / μl.

[0026] ⑵ Methylation treatment reagents (purchased from Beijing Tianmo Technology Development Co., Ltd.), including CT Conversion Reagent, M-Binding Buffer, M-Wash Buffer, M-Desulphonation Buffer, M-Elution Buffer, Zymo-Spin IC TM Column and Collection Tube.

[0027] (3) PCR MIX reaction liquid, it comprises Taq DNA polymerase, dNTPs and MgCl2; Wherein, the concentration of described Taq DNA polymerase is 50U / ml, and described dNTPs comprises dATP, dGTP, dCTP, dTTP, and their concentration is is 400 μM, and the MgCl2 concentration is 3mM;

[0028] (4) Deionized water

[0029] (5) Blank control

[0030] (6) Bottles or tubes and boxes for separately and collectively packing these reagents.

[0031] 2. The method ...

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Abstract

The invention discloses a primer and a kit for detecting methylation of MGMT (O<6>-methylguanine-DNA methyltransferase) genes, a use method of the primer and application of the primer and the kit. The primer disclosed by the invention is a specific primer designed according to sequences of promoter regions of the human MGMT genes, and the kit containing the primer can be used for rapidly and accurately detecting the methylation of the MGMT gene with high sensitivity through the combination of nest-shaped PCR (Polymerase Chain Reaction) amplification and DHPLC (Denaturing High Performance Liquid Chromatography) detection.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and relates to primers and a kit for gene detection, in particular to a primer and a kit for detecting MGMT gene methylation and a method for using the same Background technique [0002] Alkylating agent is a common mutagenic and carcinogenic substance. Many active groups on the alkylating agent can form many adducts with DNA, and it is particularly easy to form O with guanine. 6 -Methylguanine, O 6 - If methylguanine cannot be discovered and corrected in time, it may make it difficult to continue replication or cause mutations in replication during the DNA replication process, which may change the sequence of the genome and affect gene expression. o 6 -methylguanine DNA methyltransferase (O 6 -methylguanine-DNA methyltransferase, MGMT) is a DNA repair enzyme that can specifically repair cell damage caused by alkylating agents, and is an important link for mammalian cells to repair D...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6848C12Q2549/119C12Q2565/137
Inventor 刘晓峰程宇侯然朱立文梁超王绪华方国伟黄士昂陈忠
Owner 北京海思特医学检验实验室有限公司
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