Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Five-color fluorescence STR parting method for synchronously detecting 23 gene loci and special kit of method

A gene locus and locus technology, applied in the biological field, can solve the problem of waste of DNA database data resources, etc.

Pending Publication Date: 2019-07-30
BGI FORENSIC TECH (SHENZHEN) CO LTD
View PDF4 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, as the application of DNA as an identification method is becoming more and more extensive, users have higher and higher requirements for the number of gene loci, information content, amplification time, and the scope of application of test materials, etc. of the kit. As the scale of DNA database construction in my country continues to expand, the role of data comparison is becoming increasingly important. Since data comparison is based on the same STR gene locus, it is also necessary for the STR kit to be compatible with the gene locus, otherwise It will lead to the waste of some data resources in the DNA database

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
  • Five-color fluorescence STR parting method for synchronously detecting 23 gene loci and special kit of method
  • Five-color fluorescence STR parting method for synchronously detecting 23 gene loci and special kit of method
  • Five-color fluorescence STR parting method for synchronously detecting 23 gene loci and special kit of method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1, the preparation of the compound amplification system based on 23 gene loci

[0043] 1. Screening of 23 gene loci

[0044] In order to meet the requirements of the amount of information, individual recognition and compatibility of the detection of STR gene loci, the inventors of the present invention adopted five-color fluorescent labels to increase the number of accommodated loci, and at the same time increased the compatibility of the kit’s loci, making it compatible with white The basic standard allelic loci of the race, yellow race, black race and brown race. The inventors of the present invention finally screened a total of 23 gene loci including 21 autosomal STR gene loci, 1 sex identification locus AMEL and 1 indel locus Yindel. The 21 autosomal STR loci are D3S1358, D13S317, D7S820, D16S539, D8S1179, Penta D, D19S433, D5S818, D21S11, TPOX, D1S1656, D6S1043, D2S441, D12S391, D2F8, D13PO38CS, E5S01, Penta and FGA.

[0045] 2. Screening of primers ...

Embodiment 2

[0099] The type detection of the DNA standard 9948 based on the multiple amplification system of 23 gene loci prepared in Example 2 and Example 1

[0100] DNA standard 9948 (concentration: 10ng / μL) is a product of Xinhai Biotechnology Co., Ltd.

[0101] 1. Acquisition of DNA Standard Diluent

[0102] Take 1 μL of DNA standard 9948 and add 9 μL of TE buffer to obtain a dilution of DNA standard 9948 (concentration: 1 ng / μL).

[0103] 2. Take 15 μL of the complex amplification system based on 23 gene loci prepared in step 4 of Example 1, add 0.4 μL hot start Taq enzyme (including 2U), 1 μL DNA standard 9948 dilution and 8.6 μL nuclease-free water , to get the reaction system.

[0104] 3. Put the reaction system obtained in step 2 into a Bioer G-1000 thermal cycler for PCR amplification to obtain PCR amplification products. See Table 6 for the temperature cycle conditions of the Bioer G-1000 thermal cycler.

[0105] Table 6. Temperature Cycling Conditions

[0106]

[0107]...

Embodiment 3

[0117] Example 3, the typing detection of a triplet family based on the multiple amplification system of 23 gene loci prepared in Example 1

[0118] The blood samples of the triplet family (the identification result is identified) were provided by Guangdong Huada Forensic Evidence Identification Institute.

[0119] 1. Obtaining genomic DNA from family blood samples

[0120] Genomic DNA was extracted from the blood samples of triplet families by chelex-100 method.

[0121] 2. Take 15 μL of the complex amplification system based on 23 gene loci prepared in step 4 of Example 1, add 0.4 μL of hot-start Taq enzyme (including 2 U), 1 μL of genomic DNA from the family blood sample and 8.6 μL of nuclease-free water , to get the reaction system.

[0122] 3. Same as step 3 of embodiment 2.

[0123] 4. Same as step 4 of embodiment 2.

[0124] 5. Same as step 5 of embodiment 2.

[0125] 6. Same as step 6 of embodiment 2.

[0126] 7. Same as Step 7 of Example 2.

[0127] The typing ...

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 five-color fluorescence STR parting method for synchronously detecting 23 gene loci and a special kit of the method. The method comprises the following steps that a genome DNA of a to-be-detected individual is used as a template, the kit is adopted for PCR amplification, and a PCR amplification product is obtained; the PCR amplification product is taken, subjected to degeneration and then subjected to capillary electrophoresis detection, and genotypes of the 23 gene loci of the genome DNA of the to-be-detected individual are obtained; then an STR parting result of theto-be-detected individual is obtained. The kit comprises a primer pair combination, nucleotide sequences of 46 primers forming the primer pair combination are shown in the sequences 1-46 in a sequence table in sequence, and one primer in each primer pair is marked with fluorescence. The kit has high individual identification capacity and stability, is compatible with basic standard allele loci ofcaucasian, xanthoderm, melanoderm and brown race and is suitable for individual identification and affinity authentication of the Chinese nation population. The kit has high application value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a five-color fluorescent STR typing method for synchronous detection of 23 gene sites and a special kit thereof. Background technique [0002] Short tandem repeats (short tandem repeats, STRs) are a class of molecular genetic markers that widely exist in eukaryotic genomes and belong to the second generation of genetic markers. In the human genome, there is a STR site about every 6-10kb, and they account for 10% of the total human genome. These STRs usually consist of tandem repeating units of 2-6 bases. Due to the difference in repeating units and repeating times, they show large differences among people of different races and regions, which is manifested as genetic polymorphism. Compared with other genetic markers, STR genetic markers not only have a large amount of information, but also have large differences among different individuals, high polymorphism, small fragm...

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 Applications(China)
IPC IPC(8): C12N15/11
CPCC12Q1/6876C12Q1/686C12Q2600/16C12Q2563/107C12Q2537/143C12Q2565/125
Inventor 李生斌伏东科刘松娇杨日华
Owner BGI FORENSIC TECH (SHENZHEN) CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com