Forensic fluorescence composite detection kit based on 9 slow-mutation Y chromosome STR genetic markers

A Y chromosome and composite detection technology, applied in the field of forensic medicine, can solve the problems of high cost and time-consuming STR detection, and achieve the effect of peak balance, high sensitivity and consistent typing

Inactive Publication Date: 2020-03-13
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, the current detection technologies for SNP include single base extension reaction, Sanger sequencing, massively parallel sequencing and SNP chip technology, but these technologies are relatively expensive and time-consuming compared with STR detection.

Method used

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  • Forensic fluorescence composite detection kit based on 9 slow-mutation Y chromosome STR genetic markers
  • Forensic fluorescence composite detection kit based on 9 slow-mutation Y chromosome STR genetic markers
  • Forensic fluorescence composite detection kit based on 9 slow-mutation Y chromosome STR genetic markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Embodiment 1 prepares kit of the present invention

[0074] The forensic science fluorescent composite detection kit for detecting 9 slow mutation Y chromosome STR genetic markers can include the following reagents packaged separately:

[0075] a) Multiplex amplification primer mix. It is obtained by mixing the specific amplification primers shown in Table 1. The amplification primers are synthesized by Invitrogen Company. The synthesized amplification primers are prepared with ultrapure water to 100pM / μL, and then mixed according to the ratio in Table 1 to prepare Multiplex amplification primer mix. Table 5 shows the ratio of each primer when preparing 50 μL of compound amplification primer mixture.

[0076] b) Compound amplification reaction mixture. In this embodiment, the 2×QIAGEN Multiplex PCR Master Mix in the QIAGEN Multiplex PCR Kit of Qiagen Company was used.

[0077] c) Allelic Ladder. Perform single amplification on each allele of the 9 slow mutation Y c...

Embodiment 2

[0082] Example 2 Using the kit of the present invention to detect 210 irrelevant individual samples

[0083] Using the above-mentioned forensic medicine fluorescent composite detection kit based on 9 slow mutation Y chromosome STR genetic markers, 210 unrelated individual samples were tested. The unrelated individual samples here were from Hebi City, Pingdingshan City and Puyang City in Henan Province. The specific detection process is carried out as follows:

[0084] a. Use the PureLink Genomic DNA Mini Kit from Thermo Fisher Scientific to extract DNA from 210 unrelated individual blood stain samples.

[0085] The specific extraction process is as follows: ① Adjust the water bath to 55°C; ② Add 2-5 bloodstains of 2-3mm in size to a 1.5mL sterile microcentrifuge tube; ③ Add 180 μL PureLink Genomic Digestion Buffer and 20 μL proteinase K, shake and mix to ensure that the blood stains are completely immersed in the buffer; ④Incubate at 55°C for 30 minutes, and shake intermitten...

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Abstract

The invention belongs to the technical field of forensic examination, and specifically relates to a forensic fluorescence composite detection kit based on 9 slow-mutation Y chromosome STR genetic markers. Aiming at the problem that high mutation rate of a conventional Y chromosome STR is not conducive to evolutionary researches and low resolution of a Y chromosome SNP is not conducive to forensicapplication, the invention provides a forensic fluorescence composite detection kit based on 9 slow mutation Y chromosome STR genetic markers, which comprises a composite amplification primer mixtureof 9 slow mutation Y chromosome STR loci, a composite amplification reaction mixture, an allele genotyping standard substance and a standard DNA 9948. The 9 slow mutation Y chromosome STR loci are DYS593, DYS443, DYS645, DYS388, DYS531, DYS596, DYS426, DYS454 and DYS455. The kit of the invention has high sensitivity, good stability and strong repeatability, and can be effectively applied to researches in the fields of forensic family search, population history, population migration evolution and the like.

Description

technical field [0001] The invention belongs to the technical field of forensic medicine inspection, and in particular relates to a forensic medicine fluorescence composite detection kit based on 9 slow mutation Y chromosome STR genetic markers. Background technique [0002] The Y chromosome is a male-specific chromosome, belonging to the proximal centromere chromosome, and its DNA is about 60Mb in length. According to the genetic pattern, it can be divided into two regions, namely the pseudoautosomal region (PAR) located at both ends of the Y chromosome and the male-specific region (MSY). The pseudo-autochromatic region accounts for about 5% of the Y chromosome, and can recombine and exchange with the corresponding region of the X chromosome during meiosis. The male-specific region is also called the non-recombining region (NRY), which accounts for about 95% of the Y chromosome. It will not recombine with any chromosome and can only be inherited independently from the fath...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6858
CPCC12Q1/6858C12Q1/6888C12Q2600/156C12Q2600/16C12Q2531/113C12Q2563/107C12Q2537/143C12Q2565/125
Inventor 侯一平刘京王正宋凤郎敏李英碧
Owner SICHUAN UNIV
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