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Primer pair capable of detecting specificity of human mitochondrial genome

A mitochondrial genome and primer set technology, applied in the field of genomics, can solve the problems of inability to guarantee, high accuracy and high requirements

Inactive Publication Date: 2015-04-01
HANGZHOU JILUO BIOLOGICAL PHARMA CO LTD
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Problems solved by technology

[0005] Long-fragment sequencing has high requirements for samples and Taq enzymes, and high accuracy cannot be guaranteed

Method used

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  • Primer pair capable of detecting specificity of human mitochondrial genome
  • Primer pair capable of detecting specificity of human mitochondrial genome

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Embodiment Construction

[0027] Since the amplification of a large fragment of the mitochondrial genome has high requirements on sample quality and Taq enzyme, the one-time amplification of the mitochondrial genome will have a high mismatch rate. Currently the most accurate Sanger sequencing method has an effective read length of 700-800bp. The present invention uses 16 pairs of primers to amplify the fragments respectively, and then splicing them, which can improve the accuracy of the data. In the following, the 16 primer pairs of the present invention are applied to the specific detection of human mitochondrial genome to further illustrate the present invention.

[0028] 1. Select the research object: The experimental specimens come from the Shanghai Blood Bank, a total of 8 specimens.

[0029] 2. Extraction of whole genome DNA: operate according to the operating instructions of the blood genome DNA extraction kit, the kit is DNeasyBlood&TissueKit. DNA electrophoresis after extraction see figure ...

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Abstract

The invention discloses a primer pair capable of detecting specificity of a human mitochondrial genome. The primer pair capable of detecting the specificity of the human mitochondrial genome comprises primer sequences shown in SEQ ID NO.1-SEQ ID NO.32. Sixteen pairs of primers are designed, then direct sequencing is carried out on a PCR product, and finally a splicing method is carried out; compared with the long-fragment sequencing in the prior art, the PCR product of the designed primers covers the whole genome sequence of the whole mitochondrion, length of the PCR product of each pair of primers is 1200-1300bp, and a sequence obtained by two adjacent pairs of primers is overlapped for 100-300bp, and thus information of the mitochondrial genome can be accurately obtained at one time, all the mutation on the mitochondrial genome can be detected, and diagnosis of mitochondrion-related diseases can be assisted.

Description

technical field [0001] The invention belongs to the field of genomics and relates to a novel amplification primer for the whole genome sequence of human mitochondria. Background technique [0002] Mitochondria are very important organelles in eukaryotic cells, the main part of eukaryotic oxidative metabolism, and the place where carbohydrates and other substances are finally oxidized in cells to release energy. Mitochondria itself has an independent set of genome, which only divides in oocytes, so it has the characteristics of maternal inheritance, that is, it is only passed on to the next generation through women. The transmission of mtDNA mutations has a number of features. [0003] Each cell contains a large number of mitochondria. If a certain site on all the mitochondrial DNA in the cell is the same type of base, that is, all are normal genes or mutant genes, the cell is pure; if A cell is heterogeneous when both normal and mutant genes exist at a certain site on all ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6869
Inventor 李洲范嘉庚李航
Owner HANGZHOU JILUO BIOLOGICAL PHARMA CO LTD
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