26-pair PCR primer for mitochondrion sequencing and parting method based on the primer

A mitochondrial and sequencing technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as poor gene frequency distribution, inability to reflect mitochondrial sequence characteristics, and inability to achieve standardization.

Inactive Publication Date: 2008-09-24
XI AN JIAOTONG UNIV
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Problems solved by technology

[0003] (1) There is no unified scheme for the existing mitochondrial base sequence determination methods, and artificial selection is required
[0004] (2) These mitochondrial base sequences are all based on the data obtained from the determination of groups other than the Chinese population (mainly the white population). Some of the gene loci have poor gene frequency distribution in the Chinese population an

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  • 26-pair PCR primer for mitochondrion sequencing and parting method based on the primer

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Embodiment Construction

[0020] The PCR primers for mitochondrial sequencing of the present invention are shown in Table 1.

[0021] Based on the typing method of the PCR primers for mitochondrial sequencing, the present invention utilizes 26 pairs of PCR primers covering the entire length of the mitochondrial genome to amplify target fragments, perform direct sequencing detection, and perform typing.

[0022] 1. Design primer sequences

[0023] The PCR primer sequences were designed with reference to the mitochondrial standard sequence published by Genebank (http: / / www.ncbi.nlm.nih.gov), and the upstream and downstream primer sequences are shown in Table 1. Mitochondrial DNA sequence is circular, primer design strategy see figure 1 .

[0024] The 26 pairs of PCR primers for mitochondrial sequencing were selected, and the upstream and downstream primers were diluted to 100 μMOL / L as a mother solution, and 10 μl was taken out and diluted to 5 μMOL / L as a working solution.

[0025] 2. PCR amplificati...

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Abstract

The invention relates to a detection method of nucleic acid, particularly discloses a whole base sequence typing of mitochondria and a determination method thereof, specifically 26 pairs of PCR primer for mitochondria sequencing and a typing method based on the primers. The 26 pairs of PCR primer which are adopted by the invention cover the total length of mitochondria genome, wherein, primer 15-1, 15-2, 15-3, 24-1 and 24-2 are designed renewedly based on Chinese population and has the pertinence of the typing of Chinese population. The amplified fragment which corresponds to the primer 24-1 is the minimum and equals to 420bp. The amplified fragment which corresponds to the primer 22 is the maximum and equals to 1162bp. The dimensions of all PCR fragments are moderate and favorable for PCR amplification. The 26 pairs of PCR primer of the invention is utilized in the laboratory of applicant for investigating the community information of Chinese nation, and the whole sequence information of Chinese population is submitted to GENEBANK databases. The result shows that the invention can be definitely applied in the field of individual recognitions, paternity testing and gene diagnosis of the field of forensic medicine, anthropology, genetics and disease.

Description

technical field [0001] The present invention relates to a nucleic acid detection method, in particular to a typing method for the entire base sequence of mitochondria and its determination method, in particular to 26 pairs of PCR primers for mitochondrial sequencing and a typing method based on the primers. Background technique [0002] Mitochondrial (mtDNA) base typing, currently the most commonly used method is to amplify the required DNA fragments by polymerase chain reaction (PCR), directly sequence the amplified products, and carry out the analysis according to the base sequence of the fragments and the mitochondrial standard sequence Compare the results. For the direct sequencing of PCR products, PAGE combined with silver staining and automatic analysis system of fluorescent marker sequencer can be used. Research on mitochondria in China is still focused on the specific hypervariable region of mitochondria. At present, the entire base sequence of mitochondria has been...

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 李生斌刘清波赖江华张洪波杨丽
Owner XI AN JIAOTONG UNIV
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