Unlock instant, AI-driven research and patent intelligence for your innovation.

29 micro-haplotype loci, screening methods, multiplex amplification systems and applications

A compound amplification system and an amplification system technology, which are applied in compound amplification systems and application fields, can solve the problems of indistinguishable individuals with paternal genetic relationship, unable to be successfully detected and typed, and sibling pairs unable to be distinguished, etc. Achieve the effect of fast detection speed, high detection throughput, and large amount of data obtained

Active Publication Date: 2021-12-31
NANJING MEDICAL UNIV
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when the proportion of DNA from different individuals in the mixed biological specimens differs too much (less than 5%~10%), due to the influence of dominant amplification, shadow peak (stutter), random effect and other factors, the DNA component is less Few individuals can be successfully detected and typed
When Y-STR genetic markers are used, the detection ratio of male DNA components against female DNA background can be increased, but female components cannot be detected, and it is difficult to distinguish individuals with paternal genetic relationships, even with rapid mutations with high mutation rates (rapid mutating, RM) Y-STR genetic markers, about 50% of father-son pairs and 40% of sibling pairs cannot be distinguished

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • 29 micro-haplotype loci, screening methods, multiplex amplification systems and applications
  • 29 micro-haplotype loci, screening methods, multiplex amplification systems and applications
  • 29 micro-haplotype loci, screening methods, multiplex amplification systems and applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] The screening method of 29 new micro-haplotype sites of the present invention comprises the following steps:

[0059] (1) Population data obtained from the Thousand Genomes database;

[0060] (2) Obtain micro-haplotype genotypes and allele frequencies from the population data according to the screening rules, and calculate the detailed process of forensic parameters of candidate sites in different populations;

[0061] (3) Further check the sequences where the screened micro-haplotype sites are located, and exclude sites that affect subsequent sequencing typing.

[0062] In step (1), candidate micro-haplotype loci are screened from the Thousand Genomes database, such as figure 1 as shown, figure 1 It is part of the population data obtained from the Thousand Genomes database.

[0063] In step (2), the screening rule is that the sequence length is less than 50 bp, there are 4 or more alleles, and the frequency of at least 4 alleles is greater than 0.1, and the Ae value...

Embodiment 2

[0074] The composite amplification system of 29 micro-haplotype sites of the present invention, the composite amplification system comprises:

[0075] (1) The first round of PCR amplification system

[0076] The first round of PCR amplification system uses the DNA template as the target to amplify, and the primers in the system are designed according to the selected micro-haplotype sites, and the first round of PCR amplification system also includes bridging sequences ;

[0077] (2) The second round of PCR amplification system

[0078] The second-round PCR amplification system uses the dilution of the first-round amplification product as a template to perform the second-round amplification;

[0079] (3) The third round of PCR amplification system

[0080] The third-round PCR amplification system uses the second-round PCR amplification product as a template for third-round amplification, and the primers used in the third-round PCR amplification system are complementary seque...

Embodiment 3

[0111]The present invention discloses the application of the above composite amplification system in the inspection and typing of mixed biological samples, especially the application in the inspection and typing of mixed biological samples from Chinese population. The application method includes the following steps:

[0112] (1) The DNA templates extracted from the mixed biological samples were subjected to the first round of PCR amplification system, the second round of PCR amplification system, and the third round of PCR amplification in the multi-micro haplotype site composite amplification system. Amplification of the system;

[0113] (2) Perform large-scale parallel sequencing on the final amplification results of the multi-micro-haplotype site compound amplification system to obtain the average sequencing depth of each micro-haplotype site in the DNA template.

[0114] In step (2), the fastq sequence files obtained after massively parallel sequencing were analyzed using ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a 29 micro-haplotype sites, a screening method, a compound amplification system and applications. The present invention obtains population data, obtains micro-haplotype genotypes and allele frequencies therefrom, calculates the forensic medicine-related parameters of candidate sites in different populations, and then excludes sites that affect subsequent sequencing typing, 29 micro-haplotype sites were obtained; primers and amplification conditions were designed according to the 29 micro-haplotype sites, and a composite amplification system based on three rounds of PCR amplification systems was established. The DNA template extracted from the test material is amplified, and the amplification result is subjected to large-scale parallel sequencing to obtain the value of the sequencing depth of each micro-haplotype site in the DNA template, and finally achieve the typing of the mixed biological test material Purpose. The complex amplification system of the present invention is suitable for large-scale parallel sequencing technology, and has the advantages of high detection throughput, fast detection speed, large amount of data obtained, and more accurate detection results.

Description

technical field [0001] The invention belongs to forensic material evidence identification, and in particular relates to 29 micro-haplotype sites, a screening method, a compound amplification system and applications. Background technique [0002] Mixed biological samples refer to samples that are mixed with two or more biological components. Mixed biological specimens are a common type of difficult specimens in actual forensic prosecution. With the continuous advancement of scientific and technological means, more and more criminal cases have added the inspection and use of DNA in mixed biological samples. Mixed biological samples may appear in various types of cases, and most of them are sexual assault and violent injury (such as homicide, dismemberment) cases. Suspicious dander, skin swabs, or bodily fluid evidence collected by investigators at the scene often come from different individuals. Therefore, the DNA testing and determination of mixed biological samples plays ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888
Inventor 陈峰陈鹏李开黄惠结俞延芳俞尤嘉毛征生
Owner NANJING MEDICAL UNIV