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Primer group and kit for simultaneously amplifying 13 human STR gene loci and application of primer group and kit

A gene locus and primer set technology, applied in the field of molecular genetics, can solve the problems of shortening DNA molecule length, missing alleles of large fragment STR loci, and rapid detection of unfavorable cases, achieving high sensitivity and specific amplification Strong sex, the effect of improving individual recognition

Pending Publication Date: 2021-05-28
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

However, in complex and changeable natural environments, especially environmental factors such as humidity, high temperature, ultraviolet rays, exposure to sunlight, microorganisms, and strong acids can damage DNA in biological samples, cause double-strand breaks, and shorten the length of DNA molecules.
Performing routine STR typing tests on such degraded biological samples is prone to loss of alleles at the loci of large fragments of STR loci, resulting in typing failures
It greatly limits the application of STR typing technology in trace and degradable biological samples, which leads to the failure to fully reflect the value of some important biological samples in the detection of cases, which is not conducive to the rapid detection of cases

Method used

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  • Primer group and kit for simultaneously amplifying 13 human STR gene loci and application of primer group and kit
  • Primer group and kit for simultaneously amplifying 13 human STR gene loci and application of primer group and kit
  • Primer group and kit for simultaneously amplifying 13 human STR gene loci and application of primer group and kit

Examples

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Effect test

Embodiment 1

[0100] Direct multiplex amplification was performed on two hairs from suspected father and son to detect 13 STR 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:

[0101] ①Use scissors to cut the hair short at 0.5cm above the hair follicle, and put the short hair with the hair follicle into a 200μl PCR tube.

[0102] ②A 10 μl amplification system was prepared according to the standard reaction system of the present invention: 2 μl of primer mixture, 4 μl of reaction premix...

Embodiment 2

[0109] Embodiment 2: the application of kit of the present invention in forensic medicine

[0110] 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.

[0111] 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...

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Abstract

The invention provides a primer group and a kit for simultaneously amplifying 13 human STR gene loci and application of the primer group and the kit, and belongs to the technical field of molecular genetics. In the invention, the 13 STR gene loci comprise 12 autosomal STR gene loci and one sex identification STR gene locus. According to the method, 13 STR gene loci with fragments smaller than 320 bp can be amplified in one reaction at the same time, 8 core gene loci and 5 preferable gene loci specified by the Ministry of Public Security are included, meanwhile, a sex identification gene locus is further included, and the problem that detection of trace and degraded large-fragment gene loci of a detected material is lost can be effectively prevented. The 13 STR gene loci are combined with mainstream kits in the market, and more gene locus information can be obtained for trace and degraded DNA, so that the individual recognition capability is effectively improved.

Description

technical field [0001] The invention relates to the technical field of molecular genetics, in particular to a primer set, a kit and an application thereof for simultaneously amplifying 13 human STR loci. Background technique [0002] STR (short tandem repeats, short tandem repeats), also known as microsatellite sequence, is a short tandem repeat sequence widely present in human genome 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 500bp), so the requirement for template quality is lower , 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 for Southern blot analysis. In view of the above characteristics, STR typing technology has bee...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6888C12Q1/6858C12Q2600/156C12Q2600/16
Inventor 冉凌飞蒿杰刘甲乾
Owner 百特元生物科技(北京)有限公司
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