Primer group and kit for simultaneously amplifying 44 Y-STR gene loci of human and application thereof

A Y-STR, locus technology, applied in the field of molecular genetics, can solve the problems of inconvenience and time-consuming, and achieve the effects of high individual identification, high non-father exclusion rate, and strong amplification specificity

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, most of the kits currently on the market only contain 29 core and preferred loci, and the kits of each company are different in the preferred loci, which is ext

Method used

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  • Primer group and kit for simultaneously amplifying 44 Y-STR gene loci of human and application thereof
  • Primer group and kit for simultaneously amplifying 44 Y-STR gene loci of human and application thereof
  • Primer group and kit for simultaneously amplifying 44 Y-STR gene loci of human and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0125] Direct multiplex amplification was performed on two hairs from suspected father and son to detect 44 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:

[0126] 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.

[0127] ② According to the standard reaction system of the present invention, a 10 μl amplification system was prepared: 2 μl primer mixture, 4 μl reaction premix, 4 μl d...

Embodiment 2

[0136] The invention is mainly used for the construction of DNA database and identification.

[0137] The detailed part of the operation is as follows:

[0138] ① Known personnel samples are usually blood cards, which can be directly amplified. Use a puncher with a diameter of 1mm to directly punch a blood piece with a diameter of 1mm on the dry blood card and put it into a 200μl PCR tube.

[0139] ②Unknown persons are usually the samples collected at the scene of the case, and the samples are complicated. Usually, the method of extracting DNA first and then amplifying the data is used for detection and analysis. DNA was extracted by Chelex-100 and magnetic bead method. (For the extraction method, refer to Chapter 4 DNA Extraction of "Forensic DNA Analysis" by Zheng Xiufen).

[0140] ③ For the preparation and amplification of the reaction system, refer to the standard system of the present invention to prepare a 10 μl reaction system, see Table 7 below. PCR amplification w...

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Abstract

The invention provides a primer group and a kit for simultaneously amplifying 44 Y-STR gene loci of human and application thereof, and belongs to the technical field of molecular genetics. According to the present invention, the 44 Y-STR gene loci comprise 41 Y chromosome STR gene loci and 3 Y-indel gene loci, including all the gene loci of the mainstream kit in the market; according to the invention, 44 gene loci can be simultaneously amplified in one reaction, the amplification time can be shortened, and the identification efficiency and the detection material adaptability of the kit are 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 44 human Y-STR loci. 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 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 typin...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2600/156C12Q2600/16
Inventor 冉凌飞蒿杰刘甲乾赵亚楠路瑶
Owner 百特元生物科技(北京)有限公司
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