STR typing method for loca D8S1179 based on next generation sequencing

A technology of D8S1179-RP and D8S1179, applied in the field of STR typing of locus D8S1179, which can solve the problems of allele loss, limited genetic information, and difficulty in individual identification of trace or complex problems

Active Publication Date: 2016-07-13
BEIJING CENT FOR PHYSICAL & CHEM ANALYSIS
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, with the continuous improvement of DNA extraction ability and detection technology sensitivity, forensic physical evidence detection has developed from constant detection to trace or even trace detection. Problems such as heterozygote peak imbalance, allelic drop-out,...

Method used

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  • STR typing method for loca D8S1179 based on next generation sequencing
  • STR typing method for loca D8S1179 based on next generation sequencing
  • STR typing method for loca D8S1179 based on next generation sequencing

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Experimental program
Comparison scheme
Effect test

Embodiment 1D8

[0031] Embodiment 1D8S1179 Gene Specific Fragment Amplification

[0032] With the consent of the blood donor, normal blood was collected with an anticoagulant tube, and the blood collection tube was immediately placed in an ice box. All blood samples were analyzed by molecular biology method, compared with the human genome sequence reported in NCBI database, using PCR method, according to the primer design principle, using PrimerExpressVersion3 software to design primers in the conserved sequence region of D8S1179 gene:

[0033] D8S1179-FP:5'-TTTGTATTTCATGTGTATCATTCGTATC-3'

[0034] D8S1179-RP:5'ACCTATCCTGTAGATTATTTTCACTGTG-3'

[0035] 1. Use Chelex100 kit to extract DNA from blood of different samples:

[0036] Cut an appropriate amount of blood spots, put them in a centrifuge tube for 0.5 minutes, add appropriate amount of pure water, soak at room temperature, centrifuge at 13,000 rpm for 5 minutes, discard the supernatant, leave about 20 μL of liquid and test material mat...

Embodiment 2

[0063] Example 2 Establishment of a method for analyzing biological information of the D8S1179 gene by next-generation sequencing

[0064] After the library is successfully constructed, take out all the sequencing sequences containing the sample bacode in the original data, first count the length distribution, distinguish the DNA sequence of each sample according to the index sequence in the sample primer, count the number of repeats of the same DNA molecule, and find out the occurrence of SNPs, flanking sequences were retrieved and analyzed. The results shown in Table 2 can appear:

[0065] Table 2

[0066]

[0067] It can be seen that there are 12 and 13 repeats in this sample and in the second repeat, a SNP site with A mutation to G appeared.

[0068] Embodiment 3 Application experiment 1

[0069] For the existing 9 samples of the first-generation typing results, extract DNA, use the primers of the present invention, carry out the PCR method according to the method of...

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Abstract

The invention relates to the field of molecular biology and particularly discloses a specific primer pair for amplifying a gene D8S1179. The specific primer pair comprises D8S1179-FP:5'-TTTGTATTTCATGTGTACATTCGTAATTC-3' and D8S1179-RP:5'-ACCTATCCTGTAGATTATTTTCACTGATTG-3'. The invention further provides an STR typing method for a loca D8S1179 based on next generation sequencing and particularly provides a primer combination for being connected with a proper adaptor. The STR typing method is applied to the STR typing of the loca D8S1179. According to the primer pair capable of amplifying the gene D8S1179, after concentrations of different sample products subjected to PCR amplification, the equivalent mixing is carried out, and a genome library is constructed according to a BIOO RAPID DNA kit; the library is input into a computer by virtue of a next generation sequencer, data is subjected to bioinformatics analysis, and the typing results such as times of repetition of NDA molecules with same length, SNP of the loca and the difference among side wings of the loca of D8S1179 can be found.

Description

technical field [0001] The present invention relates to the field of molecular biology, in particular to a method for typing STRs based on the gene locus D8S1179 of next-generation sequencing. Background technique [0002] One of the basic tasks of forensic physical evidence is to solve the problem of individual identification. Forensic DNA analysis plays a vital role in clarifying the nature of the case and identifying the suspects, parties and victims. Forensic DNA analysis is a science and technology that uses modern DNA technology to analyze the distribution and transmission of DNA genetic markers in the population, to determine the consistency and genetic relationship of the analyzed samples, and to provide evidence for the investigation and judicial trial. Since 1985, Jeffereys and others established the first generation of typing technology - DNA fingerprinting technology, together with the later development of PCR amplified fragment length polymorphism analysis techn...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6869C12Q1/6888C12Q2600/156C12Q2600/172C12Q2525/191C12Q2535/125C12Q2563/185
Inventor 钱嘉林刘旭武会娟严江伟萨日娜
Owner BEIJING CENT FOR PHYSICAL & CHEM ANALYSIS
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