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Specific amplification primer group for simultaneously amplifying 25 human STR (short tandem repeat) gene loci, fluorescence labeling amplification kit, application and method

A fluorescent labeling and amplification primer technology, applied in the field of molecular genetics, can solve problems such as inconvenient use of public security, achieve high non-father exclusion rate, facilitate detection, and have the effects of strong specificity

Active Publication Date: 2021-09-21
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 group for simultaneously amplifying 25 human STR (short tandem repeat) gene loci, fluorescence labeling amplification kit, application and method
  • Specific amplification primer group for simultaneously amplifying 25 human STR (short tandem repeat) gene loci, fluorescence labeling amplification kit, application and method
  • Specific amplification primer group for simultaneously amplifying 25 human STR (short tandem repeat) gene loci, fluorescence labeling amplification kit, application and method

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Effect test

Embodiment 1

[0097] Kit optimization

[0098] In order to enable the performance of the present invention to meet the expected targets: simultaneous amplification of 25 loci, high sensitivity, adaptability to various types of test materials, rapid amplification and other indicators, the present invention also optimizes each part of the kit.

[0099] (1) Optimize the composition of the kit

[0100] The present invention can simultaneously amplify 25 loci fluorescent marker composite amplification kit, which includes: primer mixture, PCR premix, deionized water, positive control DNA M2 or 9948, allelic ladder 25A, molecular weight internal standard BTY -550, six-color spectrum calibration reagent (including blue, green, yellow, red, purple, orange).

[0101] Positive control DNA M2 or 9948 is an amplification standard used to test the quality of the amplification system. In the preferred solution of the present invention, all positive controls can be correctly typed, and the sensitivity is...

Embodiment 2

[0123] Application of the kit of the present invention in identification of kinship

[0124] Direct multiplex amplification was performed on two hairs from suspected father and daughter to detect 25 loci. The amplification method adopts the method of direct amplification: take a section of hair with hair follicles and put it directly into the amplification system so that the hair follicles are completely immersed in the liquid. The amplification procedure adopts the standard amplification procedure of the present invention, and the amplification instrument adopts ABI Proflex PCR instrument, genetic analyzer adopts 3500xl genetic analyzer, analysis software adopts GeneMapper-ID-X analysis software. The operation steps of this embodiment are as follows:

[0125] ①Use scissors to cut short the hair 0.5cm above the hair follicle, and put the short hair with hair follicle into a 200μL PCR tube. The hair used for augmentation must have hair follicles. If there are no hair follicle...

Embodiment 3

[0133] Application of the kit of the present invention in forensic medicine

[0134] The invention is mainly used in the field of forensic medicine for the construction of the DNA database of criminals and identification of the identity of criminal suspects.

[0135] The national forensic DNA database is a national DNA database of criminals established by the Ministry of Public Security of my country. Now the database has exceeded 50 million data, and it is the main tool for the public security system to detect cases. The database is usually built by the method of direct amplification and detection of blood samples. The identification of illegal and criminal suspects is usually complicated, and the method of amplification and detection after DNA extraction is used for detection. The invention can take into account two different inspection materials, and facilitates the construction of a national DNA database and identification of the identity of criminal suspects. The detaile...

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Abstract

The invention relates to a specific amplification primer group for simultaneously amplifying 25 human STR (short tandem repeat) gene loci, a fluorescence labeling amplification kit, application and a method, and belongs to the technical field of molecular genetics. The specific amplification primer group disclosed by the invention comprises primers with nucleotide sequences as shown in SEQ ID NO: 1-50, and 25 corresponding human STR gene loci comprise 23 autosomal STR gene loci and 2 sex identification STR gene loci. The loci corresponding to the specific amplification primer group comprise all loci required by mainstream cases in the market at present, and also comprise two gender identification loci, so that the risk of gender identification errors caused by Y chromosome deletion can be effectively prevented. and the 25 gene loci are combined, so that the method has the characteristics of high individual recognition capability and high non-father exclusion rate.

Description

technical field [0001] The invention relates to the technical field of molecular genetics, in particular to a specific amplification primer set for simultaneously amplifying 25 human STR gene loci, a fluorescent marker amplification kit, an application and a method. Background technique [0002] STR (short tandem repeats, short tandem repeats), also known as microsatellite sequences, is a short tandem repeat sequence that exists in large quantities in human genomic DNA, and the repeat unit is 2 to 6 nucleotides. Due to its high polymorphism and stability, and compared with AMP-FLP and VNTR genotyping methods, the amplified product length of STR genotyping method is much smaller (less than 500 bp), so the requirement for template quality is relatively high. Low, even degraded DNA templates can be analyzed. In addition, STR typing is applicable to DNA purified by various DNA purification methods, but the amount of DNA obtained by these purification methods is often not enough...

Claims

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Application Information

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Patent Type & Authority Applications(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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