Human mitochondrial genome methylation detection kit and application thereof

A technology of mitochondrial genome and detection kit, which is applied in the field of molecular biology and can solve problems such as low sensitivity, poor accuracy, and low throughput

Active Publication Date: 2018-11-20
HEBEI MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Aiming at the technical problems of existing detection methods such as low sensitivity or poor ac...

Method used

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  • Human mitochondrial genome methylation detection kit and application thereof
  • Human mitochondrial genome methylation detection kit and application thereof
  • Human mitochondrial genome methylation detection kit and application thereof

Examples

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Embodiment 1

[0026] The embodiment of the present invention provides a human mitochondrial genome methylation detection kit, including 37 pairs of PCR amplification primers, the nucleotide sequences of the 37 pairs of PCR amplification primers are as shown in SEQ ID NO.1~SEQ ID Shown in NO.74. The kit also includes blood extraction kit (QIAamp Blood Midi Kit, QIAGEN), DNA bisulfite conversion kit (EZ DNA Methylation-Gold Kit, ZYMO), PCR amplification kit (PCR Accessory Set, SEQUENOM) , in vitro transcription and base-specific digestion kit (MassCLEAVE Kit, SEQUENOM), PCR product purification and mass spectrometry detection kit (Spectro Arrays and Clean Resin Kit, SEQUENOM).

Embodiment 2

[0028] The embodiment of the present invention provides a method for detecting methylation of human mitochondrial genome, comprising the following steps:

[0029] a) Collect samples and extract DNA

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Abstract

The invention relates to a human mitochondrial genome methylation detection kit and application thereof, wherein the kit comprises a PCR amplification kit, and the PCR amplification kit comprises 37 pairs of PCR amplification primers, and the nucleotide sequence of the 37 pairs of PCR amplification primers is shown as SEQ ID NO. 1 to SEQ ID NO. 74. The kit can meet actual work requirements on human mitochondrial genome methylation detection in the aspects of sensibility, accuracy, stability, amplification specificity, high flux, and the like.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a human mitochondrial genome methylation detection kit and its application. Background technique [0002] DNA methylation is one of the important ways of epigenetic modification and regulation of gene expression. Mitochondria are semi-autonomous organelles with their own unique DNA (mitochondrial DNA, mtDNA). Some studies believe that methylation occurs in mitochondrial DNA and is closely related to some diseases. However, due to the sensitivity and accuracy of detection methods, some other studies believe that the degree of mitochondrial DNA methylation is extremely low, or even almost no methylation occurs. [0003] At present, there are several methods commonly used to detect DNA methylation: methylated DNA co-immunoprecipitation sequencing technology, methylation chip technology, genome-wide methylation sequencing technology, pyrosequencing and mass spectrometry. ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6858
CPCC12Q1/6858C12Q2531/113C12Q2521/525C12Q2521/327C12Q2523/125
Inventor 李淑瑾丛斌杜情情付光平
Owner HEBEI MEDICAL UNIVERSITY
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