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Kit for detecting X-STR gene seat based on next generation sequencing technology and dedicated primer combination thereof

A technology of X-STR and primer combination, applied in the field of forensic medicine

Active Publication Date: 2018-06-19
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

Illumina also launched a next-generation sequencing kit containing X-STR, namely ForenSeq TM DNA Signature Prep Kit, but it only contains 7 X-STR loci (DXS10135, DXS8378, DXS7132, DXS10074, DXS10103, HPRTB and DXS7423), and the data analysis program that comes with the kit can only target the inherent genes in the kit The analysis of the sequencing data of the seat has certain limitations

Method used

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  • Kit for detecting X-STR gene seat based on next generation sequencing technology and dedicated primer combination thereof
  • Kit for detecting X-STR gene seat based on next generation sequencing technology and dedicated primer combination thereof
  • Kit for detecting X-STR gene seat based on next generation sequencing technology and dedicated primer combination thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0125] Example 1. Preparation of a kit for detecting the X-STR locus based on next-generation sequencing technology

[0126] 1. Preparation of primer combinations

[0127] The primer set consisted of 32 primers for the detection of 16 X-STR loci. The name of each X-STR locus, the name of the primer corresponding to the amplified X-STR locus, the nucleotide sequence of the primer, and the range of allelic genotypes are shown in Table 1. The primers corresponding to each X-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.

[0128] Table 1

[0129]

[0130]

[0131] Table 2

[0132]

[0133]

[0134] 2. Preparation of kits for detecting X-STR loci based on next-generation sequencing technology

[0135] The kit for detecting the X-STR locus based on next-genera...

Embodiment 2

[0136] Example 2. Application of the kit for detecting the X-STR locus based on next-generation sequencing technology

[0137] 1. DNA sample preparation

[0138] Genomic DNA of a female oral epithelial cell sample was extracted, and then diluted with ultrapure water to obtain an aqueous solution of genomic DNA of female oral epithelial cells with a concentration of 0.425 ng / μL.

[0139] 2. Library preparation

[0140] 1. PCR amplification

[0141] Using the aqueous genomic DNA solution of female oral epithelial cells 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 PCR amplification products.

[0142] The reaction system is 20 μL, consisting of 10 μL Master Mix (product of the Physical Evidence Identification Center of the Ministry of Public Security), 4 μL primer mix, 2.5 μL genomic DNA aqueous solution of female oral epithelial cells and 3.5 μL ddH 2 O composition. In this reacti...

Embodiment 3

[0154] Example 3. Verification of the accuracy of the kit for detecting the X-STR locus based on next-generation sequencing technology

[0155] 1. DNA sample preparation

[0156] Genomic DNA of a female oral epithelial cell sample was extracted, and then diluted with ultrapure water to obtain an aqueous solution of genomic DNA of female oral epithelial cells with a concentration of 0.425 ng / μL.

[0157] Second, capillary electrophoresis detection

[0158] Take 1ng of the genomic DNA of female oral epithelial cells obtained in Step 1, and perform capillary electrophoresis detection according to the instructions of DNATyper X19 (product of the Physical Evidence Identification Center of the Ministry of Public Security), to obtain the allelic genotype of the locus. The typing results are detailed in column 2 of Table 4.

[0159] Third, next generation sequencing technology detection

[0160] The genomic DNA of the female oral epithelial cells obtained in step 1 was detected acc...

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Abstract

The invention discloses a kit for detecting X-STR gene seats based on a next generation sequencing technology and a dedicated primer combination thereof. The primer combination provided by the invention consists of 32 DNA molecules shown as sequence 1 to sequence 32 in a sequence table. Experimental results show that the kit for detecting the X-STR gene seats based on the next generation sequencing technology provided by the invention is used for detecting STR classification of 16 X-STR gene seats in genome DNAs of female oral epithelial cells, and the classification result is accurate. The kit provided by the invention has important application values.

Description

technical field [0001] The invention relates to the technical field of forensic medicine, in particular to a kit for detecting an X-STR locus based on a next-generation sequencing technology and a special primer combination thereof. Background technique [0002] In modern forensic science, DNA physical evidence plays an important role, and will become more and more important with the development of technology. At present, DNA physical evidence mainly uses capillary electrophoresis (capillary electrophoresis, CE) to detect length polymorphisms of STR loci. However, CE can only type the length of the locus, and all alleles with the same number of nucleotides are considered to be the same allele, which can easily lead to some alleles with different sequences but the same length being identified as the same allele Genes, and the sequence differences in the flanking parts cannot be detected and utilized, which seriously affect the identification of different individuals in the c...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6869C12N15/11
CPCC12Q1/6869C12Q1/6888C12Q2600/156C12Q2535/122
Inventor 季安全王乐叶健余政梁刘京武波吴浩
Owner INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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