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

A composite amplification and susceptibility technology, which is applied in the direction of material analysis, biochemical equipment and methods, and instruments by optical means, which can solve the sensitivity limitation, the number of sites in the composite amplification kit cannot be too many, and it is difficult to distinguish the test materials. genotype, etc.

Inactive Publication Date: 2008-04-09
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, otherw

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  • Multiple colour fluorescent composite amplification 11 pulmonary cancers susceptibility related SNP site kit
  • Multiple colour fluorescent composite amplification 11 pulmonary cancers susceptibility related SNP site kit
  • Multiple colour fluorescent composite amplification 11 pulmonary cancers susceptibility related SNP site kit

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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 / pl 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 disposable multicolor fluorescence combined augmentation reagent kit for 11 SNP gene loci which are related to lung cancer/munity for extracorporeal use, comprising separately packed primer mixture, DNA polymerase, buffer solution, quality control DNA specimen and allele parting normal mixture. Wherein, the primer mixture consists of 11 sharing primers marked by fluorescence and 22 length specificity primers specific to different alleles, the invention particularly consists of 2.0ml of primer mixture (therein gene locus has different primer consistency, 30nM-240nM), 1000u of DNA polymerase, 2.0ml of buffer solution, 1ml of the total amount of allele parting normal mixture, and 0.1ml (10ng/ul) of quality control DNA. Under the condition of best primer mixture ratio, the invention can rapidly and accurately obtain the genotypes of the 11 SNP points related to the lung cancer munity.

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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IPC IPC(8): G01N21/00C12Q1/68
Inventor 王瑞恒
Owner LIAONING NORMAL UNIVERSITY
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