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Gene polymorphism detection primer, probe and kit

A technology for gene polymorphism and detection reagents, which is applied in biochemical equipment and methods, microbe determination/inspection, DNA/RNA fragments, etc., can solve problems such as harsh storage conditions, improve accuracy and stability, save The effect of the thawing process

Pending Publication Date: 2020-04-10
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the storage conditions of current PCR detection reagents are usually relatively harsh, and the enzyme, reaction buffer and primers need to be stored separately, and the temperature should be around -20°C. Thaw under mild conditions

Method used

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  • Gene polymorphism detection primer, probe and kit
  • Gene polymorphism detection primer, probe and kit
  • Gene polymorphism detection primer, probe and kit

Examples

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Embodiment 1

[0057] 1. Design primers and probes for the ALDH gene, and use β-globin as an internal reference gene to design primers and probes, specifically:

[0058] ALDH2 primers are: F1:5'-CCCAGTCACCCCTTTGGTGGC-3';

[0059] R1: 5'-CAGGTCCTGAACCTCTGGC-3';

[0060] ALDH2 wild-type probe: 5'-FAM-AGGCATACACTGAAGTGA-MGB-3';

[0061] ALDH2 mutant probe: 5'-HEX-TGCAGGCATACACTAAAGTG-MGB-3'.

[0062] B-globin gene primers are: F1:5'-CTAAGGTGAAGGCTCATGGC-3',

[0063] R1: 5'-GTGCAGCTCACTCAGTGTGGC-3';

[0064] B-globin gene probe:

[0065] 5'-CY5-TGCCTTTAGTGATGGCCTGGCT-3';

[0066] 2. Preparation of reaction solution

[0067] Table 1 Concentration of reaction solution in each group

[0068]

[0069]

[0070] 3. The solution of heterozygous plasmid / pseudovirus and internal reference plasmid / pseudovirus was used as positive control. The solution of plasmid / pseudovirus containing only internal reference was used as negative control.

[0071] 4. Use the reaction solutions 1-2 recorded i...

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Abstract

The invention relates to the technical field of in-vitro diagnosis, in particular to a gene polymorphism detection primer, probe and kit. A gene polymorphism detection reagent provided by the invention comprises a reaction solution 1 and a reaction solution 2. The PCR reaction solution 1 comprises water, DNTP, DNA polymerase, UDG enzyme, Tween, DMSO, potassium acetate, NaN3, ammonium sulfate, glycerinum and Tricine; and the PCR reaction solution 2 comprises water, manganese acetate and NaN3. The reaction solution 1 can further comprise the primer and the probe. The components are reasonable incomposition, so that the reaction solutions provided by the invention can be stably stored at 2-8 DEG C, a sample can still be well detected after the reaction solutions are placed for 6 months, andunfreezing is not needed before reaction.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnosis, in particular to gene polymorphism detection primers, probes and kits. Background technique [0002] Genetic polymorphism refers to the presence of two or more genotypes at a locus in the same population, also known as genetic polymorphism. At present, human gene polymorphism is of great significance for elucidating the human body's susceptibility to disease, toxin tolerance, drug metabolism difference and the molecular mechanism of genetic diseases; polymorphism sites linked to disease-causing genes can be used as It can provide the basis for the isolation and cloning of disease-causing genes; the correlation analysis between diseases with unknown etiology and candidate gene polymorphisms can be used to assist in the screening of disease-causing susceptibility genes. Therefore, the accurate detection of gene polymorphisms plays an important role in elucidating the susceptibility and ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6858C12N15/11
CPCC12Q1/6858
Inventor 罗培李江浩段晓蒙李振红李静静杜美付光宇吴学炜苗拥军
Owner AUTOBIO DIAGNOSTICS CO LTD
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