Primer group complexly amplified by fluorescent mark for synchronously analyzing 27 genetic locus of human genomic DNA, kit and application

A technique of fluorescent labeling and multiple amplification, applied in the field of molecular biology, to achieve good allele frequency distribution, high genetic polymorphism, and increase the possibility of effect

Inactive Publication Date: 2015-07-01
AGCU SCIENTECH
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, with the increasing demand for the number of detection loci, the limitation of the number of detection loci by the five-color fluorescence technology is...

Method used

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  • Primer group complexly amplified by fluorescent mark for synchronously analyzing 27 genetic locus of human genomic DNA, kit and application
  • Primer group complexly amplified by fluorescent mark for synchronously analyzing 27 genetic locus of human genomic DNA, kit and application
  • Primer group complexly amplified by fluorescent mark for synchronously analyzing 27 genetic locus of human genomic DNA, kit and application

Examples

Experimental program
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Effect test

Embodiment 1

[0042] 1. Determination of genetic loci

[0043] Perform in-depth analysis of existing STR loci and optimize new high-discrimination loci. 对D1S1656、D6S1017、D4S2408、D8S1179、D21S11、D7S820、D13S317、D16S539、TPOX、TH01、D5S818、vWA、D18S51、FGA、D2S1338、D19S433、Penta E、D10S1248、D1S1677、Penta D、CSF1PO、D3S1358、D19S253、D12S391 、D6S1043、D6S474、D12ATA63、D22S1045、D11S4463、D1S1627、D3S4529、D2S441、D17S1301、D1GATA113、D18S853、D20S482、D14S1434、D5S2500、D2S1776、D10S1435和D9S1122等40多个STR基因座在中国人群的遗传多态性开展 Research. Genotype detection was performed on more than 5,000 individuals, and data such as individual recognition ability (DP), heterozygosity (H), and non-parent exclusion rate (PE) were calculated according to the distribution frequency of alleles at each locus, indicating that the 24 STR genes Among the loci, except for the TH01 and TPOX loci, the DP values ​​of the other loci are close to 0.9, the H values ​​are greater than 0.7, and the PE values ​​are mostly above 0.5, which shows that they have go...

Embodiment 2

[0071] Example 2 Application of 27 loci fluorescent marker multiple amplification test system for triplet paternity identification

[0072]1. Collect blood samples in paternity testing cases: the samples in this experiment are provided by XX paternity testing agency. DNA extraction Use the Chelex-100 method to extract genomic DNA from 3 whole blood samples: take 0.5-5 μl of whole blood and place it in a sterilized 1.5ml centrifuge tube, add 1ml sdH2O to the tube, shake for a few seconds; place at room temperature for 10 minutes Afterwards, shake for a few seconds, centrifuge at 12,000rpm for 3 minutes, discard the supernatant, keep enough supernatant to cover the precipitate, do not stir the precipitate; add 200μl 5% Chelex-100, shake for a few seconds; incubate at 56°C for 30 minutes, shake A few seconds; boiling water bath for 10 minutes, shake for a few seconds; centrifuge at 12,000rpm for 5 minutes, the supernatant is the extracted genomic DNA. Genomic DNA was extracted w...

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Abstract

The invention relates to a primer group for fluorescence labeling compound-amplification used for synchronously analyzing 27 genetic loci of human genomic DNA, a kit and application thereof the kit and belongs to the technical field of molecular biology. 27 genetic loci are divided into five groups and totally relate to fluorescence labels of six colors; a fluorescence labeling compound amplification system disclosed by the invention is high in sensitivity and capable of detecting all 27 genetic loci when the DNA template amount is 0.12ng. The system is suitable for directly amplifying blood filter paper and FTA card acquired samples, and can be applied to individual recognition and paternity test.

Description

technical field [0001] The invention relates to a primer set, kit and application for simultaneous analysis of 27 loci of human genome DNA for fluorescent labeling compound amplification, belonging to the technical field of molecular biology, and the system can be applied to individual identification and paternity identification. Background technique [0002] Short tandem repeat locus (Short Tandem Repeat, STR) is a commonly used genetic marker. Compared with other genetic markers, STR markers have small fragments and are easy to amplify, and are more suitable for trace and degraded samples. The amplification conditions of each locus are similar and can be multiplexed, so it is fast, efficient, and accurate. , sensitivity, large amount of information and so on. Especially in the establishment of DNA database, compared with the previous RFLP and other technologies, STR multiplex amplification technology has great advantages. Therefore, the US FBI has conducted a lot of rese...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q1/6853C12Q2600/16C12Q2525/151
Inventor 郑卫国王艳芳齐丽媛张雷卢文翔郭育林葛海鹏
Owner AGCU SCIENTECH
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