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

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...

AI Technical Summary

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 and cannot reflect the Chinese population. Mitochondrial sequence characteristics of

[0005] (3) Mitochondrial assay methods published abroad are still being improved, and the standardization of results cannot be achieved among different laboratories

[0006] The above shortcomings limit the application of mitochondrial genetic markers in China, which is not conducive to further promotion at the grassroots level

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