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13 SNP sites multiple color fluorescent composite amplification reagent kit used for personal recognition

A compound amplification and kit technology, which is applied in the field of forensic personal identification, can solve the problems such as the limited number of sites in the compound amplification kit, the limitation of sensitivity, and the difficulty in distinguishing the genotype of the test material.

Inactive Publication Date: 2009-09-23
LIAONING NORMAL UNIVERSITY
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Problems solved by technology

However, since the obtained result is only a single color, the number of sites in the multiplex amplification kit should not be too many. Generally, the number of sites for one multiplex amplification cannot exceed 4, otherwise the amplification products of different sites will overlap with each other. Difficult to distinguish genotypes of specimens
In addition, the obtained results are observed with the naked eye, so its sensitivity is limited

Method used

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  • 13 SNP sites multiple color fluorescent composite amplification reagent kit used for personal recognition
  • 13 SNP sites multiple color fluorescent composite amplification reagent kit used for personal recognition
  • 13 SNP sites multiple color fluorescent composite amplification reagent kit used for personal recognition

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Embodiment Construction

[0013] This kit is composed of 13 SNP site primer mixtures, DNA polymerase, reaction buffer (including MgC12), 13 SNP site allele typing standards and quality control DNA, which are packaged separately. 2.0ml of primer mixture for 13 SNP loci (the concentration of primers for different loci is different, from 30nM-240nM, the amount used in each 20ul compound PCR reaction system is 2ul); DNA polymerase 1000u (5u / p1 for each 20ul The amount used in the compound PCR reaction system is 1u); buffer 2.0ml (10 times concentrated buffer solution, the amount used in each 20ul compound PCR reaction system is 2ul); the total amount of allele typing standard mixture 1ml (40nM, for 1000 detections), the allele typing standard mixture contains the same amount of allele typing standard at each locus; quality control DNA 0.1ml (10ng / ul).

[0014] The technical core of fluorescent multiplex amplification mainly lies in the position of primer sequence and fluorescent label and the concentration...

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Abstract

The invention relates to a kit for in vitro one-time multi-color fluorescence composite amplification of 13 SNP loci with high individual recognition ability, which is composed of a primer mixture, DNA polymerase, buffer, and quality control DNA packaged separately. Sample and allelic ladder mixtures were constructed. The primer mix consisted of 13 fluorescently labeled shared primers and 26 length-specific primers for different alleles. The genotypes of 13 SNP sites with high personal recognition ability can be obtained quickly and accurately by using this kit, and the genotypes can be detected and typed by an automatic fluorescent sequencer to achieve a high level of automation. The kit of the present invention has great advantages and potentials for forensic personal identification.

Description

technical field [0001] The invention relates to a kit for in vitro one-time composite amplification of 13 SNP gene loci for personal identification, which belongs to the invention in the field of forensic personal identification. Background technique [0002] In 1996, E. Lander of the United States first proposed "single nucleotide polymorphism" (single nucleotide polymorphism, SNP), which is called "the third generation genetic marker". SNP refers to a DNA sequence polymorphism caused by a single nucleotide variation at the chromosome genome level, and the frequency of the least one allele in the population is not less than 1%. It includes single-base conversions, transversions, insertions, and deletions. SNP has the following characteristics: First, SNP sites are very abundant. Distributed almost throughout the genome, there is approximately 1 SNP every 1000 bp in the genome, and the total number is about 3 million. Secondly, the mutation rate of SNP is low, the mutatio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/00C12Q1/68
Inventor 王瑞恒
Owner LIAONING NORMAL UNIVERSITY
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