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Specific amplification primer set for simultaneously amplifying 25 human str loci, fluorescent label amplification kit, application and method

A fluorescent labeling and amplification primer technology, applied in the field of molecular genetics, can solve the problem of inconvenient use of public security, achieve the effects of high non-father exclusion rate, facilitate detection, and high individual identification.

Active Publication Date: 2021-11-09
百特元生物科技(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, on the market, the kits of each company on the preferred loci are different, which is extremely inconvenient in the use process, and in addition to the increased demand for the gene loci, the public security users are mainly concerned about the amplification time and the types of applicable test materials. , sensitivity, etc. have also had higher and higher requirements.

Method used

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  • Specific amplification primer set for simultaneously amplifying 25 human str loci, fluorescent label amplification kit, application and method
  • Specific amplification primer set for simultaneously amplifying 25 human str loci, fluorescent label amplification kit, application and method
  • Specific amplification primer set for simultaneously amplifying 25 human str loci, fluorescent label amplification kit, application and method

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Experimental program
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Effect test

Embodiment 1

[0097] Optimization kit

[0098] In order to enable the present invention to achieve the desired performance indicators: simultaneous amplification of 25 loci, high sensitivity, adapted to various types of material being tested, rapid amplification of other indicators, the various parts of the kit of the present invention are further optimized.

[0099] (1) the composition of the kit to optimize

[0100] The present invention can be amplified 25 loci simultaneously fluorescently labeled complex amplification kit, comprising: a mixture of primer, the PCR premix solution, deionized water, or the positive control DNA M2 9948, allelic ladder 25A, a molecular weight standard BTY -550, six-color spectral calibration reagent (including blue, green, yellow, red, purple, orange).

[0101] The positive control DNA M2 or 9948, the standard amplification, amplification system used to test the quality is good or bad. In a preferred embodiment of the present invention, the positive control can ...

Embodiment 2

[0123] Application of the kit of the invention in the identification of genetic relationship

[0124] Two of the hair from the direct daughter suspected multiplex amplification, detection 25 loci. Direct Amplification Methods Amplification of: taking section having hair follicles, the amplification system directly into the hair follicle is completely immersed within a liquid, the present invention is amplified using standard procedures amplification procedure, using amplification instrument ABI Proflex PCR, using genetic analyzer 3500xl genetic analyzer software using GeneMapper-ID-X software. Example embodiment of the present procedure is as follows:

[0125] ① 0.5cm above the cut at the hair follicles with scissors, to cut into the hair follicle with 200μL PCR tubes. Must be used to amplify the hair follicle, the hair follicle is unavailable if not, can not obtain the correct type.

[0126] ② formulated according to standard reaction system of the present invention 10μL amplific...

Embodiment 3

[0133] Application kit of the invention in forensic among

[0134] The present invention is mainly used in the field of forensic building DNA database of criminal offenders and identification of criminal suspects identity crime.

[0135] National forensic DNA database is a national DNA database of criminal offenders construction of my country's Ministry of Public Security, the database now exceeded 50 million data, the public security system is the main tool of detection of cases. Database commonly used blood samples were amplified detection method for testing after direct amplification method of building a database to detect illegal criminal suspects identity identification of the work in the material being tested is usually complex seized material used to extract DNA. The invention may take into account two different material being tested, to facilitate the construction of the national DNA database and identification of criminal suspects identity crime. Detailed part of the oper...

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Abstract

The invention relates to a specific amplification primer set for simultaneously amplifying 25 human STR gene loci, a fluorescent marker amplification kit, an application and a method, and belongs to the technical field of molecular genetics. The specific amplification primer set of the present invention includes primers with nucleotide sequences as shown in SEQ ID NO: 1-50, and the corresponding human 25 STR loci include 23 autosomal STR loci and 2 sex identification STRs loci. The loci corresponding to the specific amplification primer set of the present invention include all the loci required by mainstream cases currently on the market, and also include 2 sex identification loci, which can effectively prevent sex identification caused by Y chromosome deletion Risk of error; 25 loci combined, characterized by high individual discrimination and high non-parent exclusion.

Description

Technical field [0001] The present invention relates to the field of molecular genetics, and more particularly to simultaneous amplification of the specific amplification primer group, fluorescent label amplification kit and application and method of simultaneous amplification of 25 STR locus. Background technique [0002] Str (Short Tandem Repeats, short series repeating sequence), also known as the microsatellite sequence, is a short series of repeating sequences existing in human genome DNA, and the repeating unit is 2 to 6 nucleotides. Due to high polymorphism and stability, the production of the product amplified by the STR gene-based method is much larger (less than 500 bp), so that the template quality is required Low, even if the degraded DNA template can also be analyzed. Further, the STR classification applies to DNA purified by a plurality of DNA purification methods, and the amount of DNA obtained by these purification methods is often not sufficient for Southern blot...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6888C12Q1/6858C12Q2600/156C12Q2531/113C12Q2525/151C12Q2565/125C12Q2563/107
Inventor 冉凌飞蒿杰刘甲乾路瑶
Owner 百特元生物科技(北京)有限公司
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