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Multiple colour fluorescent composite amplification kit for 11 pulmonary cancers susceptibility related SNP sites

A composite amplification and kit technology, which is applied in the direction of material analysis, measurement device, instrument, etc. by optical means, can solve the problem that the number of sites in the composite amplification kit cannot be too many, the sensitivity is limited, and it is difficult to distinguish the genotype of the test material. And other issues

Inactive Publication Date: 2009-08-26
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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  • Multiple colour fluorescent composite amplification kit for 11 pulmonary cancers susceptibility related SNP sites
  • Multiple colour fluorescent composite amplification kit for 11 pulmonary cancers susceptibility related SNP sites
  • Multiple colour fluorescent composite amplification kit for 11 pulmonary cancers susceptibility related SNP sites

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Embodiment Construction

[0016] This kit is composed of 11 SNP site primer mixtures, DNA polymerase, reaction buffer (containing MgCl2), and 11 SNP site allelic typing standards and quality control DNA that are packaged separately. 2.0ml of primer mixture for 11 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).

[0017] The technical core of fluorescent multiplex amplification mainly lies in the position of primer sequence and fluorescent label and the concentra...

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Abstract

The invention relates to a kit for in vitro one-time multi-color fluorescence composite amplification of 11 SNP loci related to lung cancer susceptibility, which is composed of a separately packaged primer mixture, DNA polymerase, buffer, quality control DNA sample and Allelic ladder mixture composition. The primer mix consists of 11 fluorescently labeled shared primers and 22 length-specific primers for different alleles. The specific composition is: 2.0ml of primer mix (the concentration of primers for different loci varies from 30nM to 240nM); Enzyme 1000u; buffer 2.0ml; total amount of allele typing standard mixture 1ml; quality control DNA 0.1ml (10ng / ul). The present invention can quickly and accurately obtain the genotypes of 11 SNP sites related to the susceptibility of lung cancer under the optimal primer ratio condition.

Description

technical field [0001] The invention relates to a kit for in vitro one-time composite amplification of 11 SNP loci related to lung cancer susceptibility on chromosomes, belonging to the invention in the field of lung cancer susceptibility detection. Background technique [0002] With the completion of the precise sequence map of the Human Genome Project, the cloning and identification of functional genes and the research on the diversity of human genome have become the next technological commanding heights. Restriction fragment length polymorphisms (restriction fragment length polymorphism RFLP) and microsatellite polymorphisms (microsatellite polymorphisms) are used as the first and second generation genetic markers, in the construction of physical maps and genetic maps, and the splicing and assembly of sequence base maps. Played a decisive role in the process. [0003] In 1996, E. Lander of the United States first proposed "single nucleotide polymorphism" (single nucleoti...

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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