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Method for quickly detecting mitochondrial gene mutation or deficiency in high-flux mode

A detection method, mitochondrial technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of inability to process a large number of samples at the same time, inability to apply clinical diagnosis on a large scale, low detection sensitivity, etc., to achieve enrichment of clinical Database, detection results are accurate and reliable, and the effect of high degree of automation

Active Publication Date: 2018-06-29
广州海思医疗科技有限公司
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Problems solved by technology

However, Sanger sequencing also has certain defects, such as high sequencing cost, high background, low detection sensitivity, inability to process a large number of samples at the same time, and inability to be applied to clinical diagnosis on a large scale.

Method used

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  • Method for quickly detecting mitochondrial gene mutation or deficiency in high-flux mode
  • Method for quickly detecting mitochondrial gene mutation or deficiency in high-flux mode
  • Method for quickly detecting mitochondrial gene mutation or deficiency in high-flux mode

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Embodiment

[0020] Example A high-throughput method for rapid detection of mitochondrial gene mutations or deletions

[0021] The method for high-throughput rapid detection of mitochondrial gene mutation or deletion of the present invention, the specific steps of the experimental operation are as follows:

[0022] 1. Extract human whole genome DNA samples, and extract DNA in strict accordance with the instructions of the Human Whole Genome DNA Extraction Kit (Lifetechnologies). The specific content is as follows:

[0023] Main instruments:

[0024]

[0025] Main reagents:

[0026]

[0027] Specific steps:

[0028] 1) Take an appropriate amount of tissue and grind it thoroughly in a liquid nitrogen environment, then transfer it to a 1.5mL centrifuge tube, add 1mL Trizol, and mix thoroughly immediately;

[0029] 2) Place the mixed tissue at room temperature for 10 minutes to fully lyse;

[0030] 3) Add 200 μL of chloroform, shake and mix well, centrifuge at 4°C, 12000rpm for 10mi...

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Abstract

The invention belongs to the field of gene detection, and relates to a method for quickly detecting mitochondrial gene mutation or deficiency in a high-flux mode. The method is characterized in that after DNA of human whole genome is extracted, a specific primer is utilized for amplifying mitochondria DNA, interrupt response is performed after an amplified product is purified; after being recycled, a target fragment is modified and connected with a sequencing joint to form a DNA library; and data are sequenced and analyzed. The method can be used for quickly, comprehensively and simultaneouslydetecting information such as mutation and deficiency of the whole mitochondria DNA of a human body, has the characteristics of being time-saving and labor-saving, and strong in specificity, can timely find existence of mitochondria disease, and enriches a database of the mitochondria disease, so that basis can be provided for clinical diagnosis.

Description

technical field [0001] The invention belongs to the field of gene detection, and relates to a method for rapidly detecting mitochondrial gene mutation or deletion with high throughput. Background technique [0002] Mitochondria is an important organelle in eukaryotic cells, which provides a place for oxidative phosphorylation of cells, and plays a vital role in cell energy metabolism, cell apoptosis, and maintaining the balance of calcium and iron ions in cells. Human mitochondrial DNA (mitochondrialDNA, mtDNA) exists in the cytoplasmic mitochondria, with a full length of 16569bp. It has a double-strand closed circular structure and is divided into two parts, the coding region and the non-coding region. There are 37 genes in the coding region, including 22 encoding transfer ribonucleic acid (tRNA) and 2 encoding ribosomal ribonucleic acid (12S and 16SrRNA). ), 13 encoded polypeptides. The mtDNA genes are closely arranged, and there are no introns between the sequences. Co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806C12Q1/6869
CPCC12Q1/6806C12Q1/6869C12Q2531/113C12Q2525/191C12Q2535/122
Inventor 周煌凯钟诗龙邓美英程祖福姚啟聪徐毓璇曾川川
Owner 广州海思医疗科技有限公司
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