Method for identifying human-derived genetic relationship through two-nucleotide repeated microsatellite

A kinship, two nucleotide technology, applied in the field of DNA fingerprinting, can solve the problems of the complexity of allele classification and high mutation rate, and achieve the effect of improving identification accuracy, low mutation rate, and avoiding the interference of slip peaks.

Active Publication Date: 2017-03-22
INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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Problems solved by technology

However, there are still deficiencies when using 4-base repeat STR for genetic identification, mainly because the mutation rate is high, and some 4-base repeat STR mutation rates can reach 1-8×10 -3 / Replacement is as much as 4 times that of 2-base repeat STR, and some 4-base STRs do not show repeat differences in the core sequence, and occasionally there are single or 2 base insertions, which gives it a Classification of alleles introduces complexity

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  • Method for identifying human-derived genetic relationship through two-nucleotide repeated microsatellite
  • Method for identifying human-derived genetic relationship through two-nucleotide repeated microsatellite
  • Method for identifying human-derived genetic relationship through two-nucleotide repeated microsatellite

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[0029] Example: See Table 2 for the STR, primers, primer modification, primer final concentration, annealing temperature, final dilution factor, PCR grouping and capillary electrophoresis grouping.

[0030] The DNA extracted from the biological sample required by the method of the present invention is less, about 50ng is enough. After commercially synthesizing the primers in Table 2, these 30 pairs of primers were diluted and paired with the forward and reverse primers according to the PCR primers, and the concentration was 20 times the final concentration of the primers in Table 2, so that multiple STRs could be detected simultaneously. PCR amplification reduces workload; for example, for D1S2757, add its forward (F direction) and reverse (R direction) primers to TE solution, so that the concentration of both primers is 6.4 μM (pmol / ul). According to the experimental determination, the primers that will not interfere with each other are put together as a group, and the primer...

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Abstract

The invention discloses a method for identifying the human-derived genetic relationship through a two-nucleotide repeated microsatellite. The method comprises the step that through primer design, primer modification, PCR amplification and capillary electrophoresis detection, the human-derived genetic relationship is identified according to capillary electrophoresis detection results. According to the method, the number of adopted sites is large, and the identification accuracy is high; compared with traditional tetranucleotide repeated STR, the two-nucleotide repeated STR is lower in mutation rate, and the identification reliability is improved; the primer for amplification is modified, so that the identification results are more clear and obvious; and the method can be used for investigation of crime, paternity test or human-derived cell identification.

Description

technical field [0001] The invention relates to a DNA fingerprint technology for detecting the identity of biological samples of human origin, that is, the identification of human genetic relationship, in particular to a method for identifying human genetic relationship by using two nucleotide repeated microsatellites. Background technique [0002] Short tandem repeats (Short tandem repeats, STRs) or microsatellites (Microsatellites) are considered second-generation genetic markers. Microsatellites are named after the early centrifugation of genomic DNA and found that they are distributed like satellites around the main band of DNA centrifugation. Microsatellites are a series of tandem sequences consisting of 2 to 5 base sequences repeated 5 to 50 times, widely distributed in the human genome, and the mutation rate of microsatellites (about 5×10-4) is much higher than that of other positions The mutation rate of DNA (the point mutation rate on DNA is about 5×10-7). The mut...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q1/6888C12Q2600/156C12Q2525/151
Inventor 孙浩黄小琴杨昭庆林克勤黄铠马绍辉褚嘉祐
Owner INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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