Multicolor fluorescence composite detection method and kit of 44 SNPs (Single Nucleotide Polymorphism) loci

A composite detection and kit technology, applied in the field of forensic individual identification, can solve problems such as unsuitable for micro-quantity test materials

Active Publication Date: 2010-12-22
HEBEI MEDICAL UNIVERSITY
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Problems solved by technology

However, in the analysis of highly degraded samples, the DNA fragments analyzed by the miniSTR locus range from 80 to 150 bp. For highly degraded DNA <100 bp, the miniSTR locus analysis technology still has limitations.
Moreover, due to the limitation of the length of the amplified fragment and the type of fluorescent dye, only a small number of miniSTR loci can be amplified simultaneously in a multiplex amplifica...

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  • Multicolor fluorescence composite detection method and kit of 44 SNPs (Single Nucleotide Polymorphism) loci
  • Multicolor fluorescence composite detection method and kit of 44 SNPs (Single Nucleotide Polymorphism) loci
  • Multicolor fluorescence composite detection method and kit of 44 SNPs (Single Nucleotide Polymorphism) loci

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Embodiment Construction

[0027] 44 SNP site multi-color fluorescent composite detection kits for individual identification, including a) amplification system: PCR buffer solution, MgCl 2 , dNTPs, PCR primers, Taq DNA polymerase, template DNA, b) amplification product purification reagents: exonuclease Ⅰ and its buffer solution, shrimp alkaline phosphatase and its buffer solution, c) extension reaction reagent: extension primer and SNaPshot reaction mixture, d) Test reagents: Hi-Di formamide and GeneScan TMSize Standards-120LIZ, the PCR primers and extension primers are composed of primers for 44 SNP gene loci, and the rs numbers of the 44 SNP gene loci in the SNP database in NCBI are rs1109037, rs3780962, rs987640, rs9951171, rs430046 respectively ,rs338882,rs2342747,rs10092491,rs1821380,rs321198,rs7041158,rs13218440,rs560681,rs445251,rs8078417,rs10488710,rs7520386,rs214955,rs4530059,rs13182883,rs722290,rs6811238,rs279844,rs9905977,rs6955448,rs4288409,rs315791,rs740598,rs2272998,rs7205345 , rs1077376...

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Abstract

The invention discloses a multicolor fluorescence composite detection kit and detection method of 44 SNPs (Single Nucleotide Polymorphism) loci, which is used for individual recognition. The kit comprises a PCR (Polymerase Chain Reaction) primer, a DNA polymerase, a PCR buffer solution, an exonuclease I, a shrimp alkaline phosphatase, an extended primer and a SNaPshot reaction mixed solution; thePCR primer and the extended primer consist of primers containing 44 SNP gene loci and respectively aim to DNA fragments containing the 44 SNP loci. By adopting the invention, the genotyping of the 44SNP loci can be obtained quickly and accurately, and the individual recognition is carried out on human biological samples through detection and analysis of a fluorescence automatic typing apparatus.

Description

technical field [0001] The invention relates to forensic individual identification, in particular to the genotyping of SNP sites, in particular to a method and kit for multicolor fluorescence composite detection of 44 SNP sites for forensic individual identification. Background technique [0002] In the work of forensic medical examination, due to the influence of high temperature, humidity, sun exposure, microorganisms and other factors, the DNA molecules in biological samples are often damaged, broken, the molecules become smaller, and large fragments are lost, making it impossible to perform effective STR typing. Make the detection of the case limited to the difficult situation. At present, in the forensic DNA analysis, the techniques to solve the DNA sample typing of highly degraded samples mainly focus on two aspects: one is the miniSTR (small fragment short tandem repeat) analysis technique. This technology shortens the length of PCR product fragments by designing the...

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Application Information

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IPC IPC(8): C12Q1/68G01N21/64
Inventor 丛斌李淑瑾娄春光
Owner HEBEI MEDICAL UNIVERSITY
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