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Kit for detecting Y-STR gene locus based on next-generation sequencing technology and special primer combination thereof

A technology of Y-STR and primer combination, which is applied in the field of forensic science and can solve the problems of expensive kits

Active Publication Date: 2018-06-22
INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above kits are expensive, and the supporting data analysis programs can only analyze the sequencing data of the inherent loci in the respective kits, which has certain limitations

Method used

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  • Kit for detecting Y-STR gene locus based on next-generation sequencing technology and special primer combination thereof
  • Kit for detecting Y-STR gene locus based on next-generation sequencing technology and special primer combination thereof
  • Kit for detecting Y-STR gene locus based on next-generation sequencing technology and special primer combination thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0137] Example 1. Preparation of a kit for detecting the Y-STR locus based on next-generation sequencing technology

[0138] 1. Preparation of primer combinations

[0139] The primer set consisted of 38 primers for the detection of 19 Y-STR loci. The name of each Y-STR locus, the name of the primer corresponding to the amplified Y-STR locus, and the nucleotide sequence of the primer are detailed in Table 1. The primers corresponding to each Y-STR locus are the PCR amplification products in the hg19 human reference genome (for information on the hg19 human genome, see the website http: / / hgdownload.soe.ucsc.edu / goldenPath / hg19 / bigZips / hg19.2bit) See Table 2 for the length and nucleotide sequence.

[0140] Table 1

[0141]

[0142]

[0143] Table 2

[0144]

[0145]

[0146] 2. Preparation of kits for detecting Y-STR loci based on next-generation sequencing technology

[0147] The kit for detecting the Y-STR locus based on next-generation sequencing technology in...

Embodiment 2

[0148] Example 2. Application of the kit for detecting the Y-STR locus based on next-generation sequencing technology

[0149] 1. DNA sample preparation

[0150] Take the standard 2800M and dilute it with ultrapure water to obtain a standard 2800M aqueous solution with a concentration of 1ng / μL.

[0151] 2. Library preparation

[0152] 1. PCR amplification

[0153] Using the standard 2800M aqueous solution obtained in step 1 as a template and the primer mixture prepared in step 2 of Example 1 as primers, PCR amplification was performed to obtain a PCR amplification product.

[0154] The reaction system is 20 μL, which consists of 10 μL Master Mix (product of the Physical Evidence Identification Center of the Ministry of Public Security), 9 μL primer mix and 1 μL standard 2800M aqueous solution. In this reaction system, primer 3, primer 4, primer 11, primer 12, primer 13, primer 14, primer 15, primer 16, primer 17, primer 18, primer 19, primer 20, primer 21, primer 22, prime...

Embodiment 3

[0166] Example 3, Verification of the accuracy of the kit for detecting the Y-STR locus based on next-generation sequencing technology

[0167] Take 1ng standard product 2800M, according to DNATyper TM Capillary electrophoresis detection is performed according to the instructions of Y26 (product of the Physical Evidence Identification Center of the Ministry of Public Security), and the allelic genotype of the locus is obtained. The typing results are detailed in column 2 of Table 4.

[0168] The standard product 2800M was detected according to the steps in Example 2 to obtain the allelic genotype of the locus. The typing results are detailed in column 3 of Table 4.

[0169] The results show that the test kit for detecting the Y-STR locus based on the next-generation sequencing technology prepared in Example 1 is compatible with the DNATyper TM The genotyping results of the overlapping locus of Y26 in the standard 2800M samples are completely consistent.

[0170] Table 4

...

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Abstract

The invention discloses a kit for detecting a Y-STR gene locus based on a next-generation sequencing technology and a special primer combination thereof. The primer combination provided by the invention is composed of 38 types of DNA (Deoxyribonucleic Acid) molecules shown as a sequence 1 to a sequence 38 in a sequence table. An experiment proves that the kit for detecting the Y-STR gene locus based on the next-generation sequencing technology, provided by the invention, is used for detecting STR typing of 19 Y-STR gene loci in a standard product 2800M and a typing result is accurate. The kitprovided by the invention has important application value.

Description

technical field [0001] The invention relates to the technical field of forensic medicine, in particular to a kit for detecting Y-STR loci based on next-generation sequencing technology and a special primer combination thereof. Background technique [0002] Human Y-chromosome short tandem repeats (Y-chromosome short tandem repeats, Y-STR) refer to the short tandem repeat fragments existing in the non-recombination region on the Y chromosome. The Y-STR locus is one of the most widely used genetic markers at present. Its repeating unit is about 2-6bp. It has the biological characteristics of paternal inheritance and can be stably inherited from father to son. Y-STR detection technology is often used in forensic science for paternity identification of patrilineal families, determination of the source of unknown male corpses, exclusion of male criminal suspects, and mixed spot testing where autosomal STR detection cannot distinguish genetic information. [0003] At present, capi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6869
CPCC12Q1/6869C12Q1/6888C12Q2600/156C12Q2531/113C12Q2535/122
Inventor 叶健王乐季安全宋娇娇陈曼武波
Owner INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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