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Primer set for amplifying genes related to exercise ability and detection kit

A technology of motility and primer sets, which is applied in the determination/inspection of microorganisms, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problems of incomplete detection of genes, long time consumption, and large input of DNA templates.

Inactive Publication Date: 2019-03-15
BEIJING BIOMASION TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the more comprehensive one is the invention patent (CN201711219812.5), which covers five sports-related gene single nucleotide polymorphism sites. This patent discloses a primer set for detecting sports gene SNP. The method is capillary electrophoresis, but the detection of genes is not comprehensive, the input of DNA template is large, and it takes a long time
Invention patent (CN201310115932.6) discloses a molecular biology method for predicting the jumping potential of excellent ice and snow athletes. This method only detects ACTN3 and ACE2 genes, and the detection of genes is not comprehensive. When the number of sites increases, the operation is complicated

Method used

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  • Primer set for amplifying genes related to exercise ability and detection kit
  • Primer set for amplifying genes related to exercise ability and detection kit
  • Primer set for amplifying genes related to exercise ability and detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Oral swab DNA extraction: saliva DNA extraction is carried out by using Biotech magnetic bead method saliva and swab DNA automatic extraction kit. Add 30 μL Proteinase K to the oral swab, bathe in water at 56°C for 30 minutes, and mix well by inverting. Tear off the plastic film of the 96-well deep-well plate, add 350 μL of the incubated sample to the first column and the seventh column respectively, then add 10 μL of nucleic acid sedimentation aid and 300 μL of isopropanol, put it into the nucleic acid extractor, insert the stirring rod into the card For the tank, set the temperature as follows: cracking temperature 80°C, elution temperature 65°C. After the instrument runs, collect the DNA solution, check the quality, and store it at 4°C.

Embodiment 2

[0038] PCR amplification: the DNA of the sample to be tested extracted in Example 1 was respectively added to a 384-well plate for multiplex PCR amplification. Add the reaction system (4 μL) according to the multiplex PCR amplification into each reaction well, the reaction system: template DNA 2 μL, primer mix 0.67 μL, 10×Buffer 0.33 μL, MgCl2 (25mM) 0.27 μL, dNTP (25mM) 0.07 μL, ddH2O 0.53 μL, PCR enzyme (5U / μL) 0.13 μL. After the reaction system is prepared, seal it tightly with PCR parafilm to prevent sample evaporation, shake and mix, and centrifuge; place the sealed 384-well plate on an ABI9700 PCR instrument for reaction, reaction conditions: pre-denaturation at 95°C for 2min; (95°C for 30s, 56 °C for 30s, 72 °C for 1 min) for 45 cycles; 72 °C for 5 min, and 4 °C for 45 cycles; the PCR amplification product was obtained and centrifuged for later use.

Embodiment 3

[0040] SAP digestion: use the Sequenom platform supporting reagents and operation steps to complete the SNP site typing results of the samples to be tested. Add the PCR amplification product obtained in Example 2 into each reaction well according to the reaction system (1.33 μL) digested by SAP, the reaction system: SAP×Buffer 0.11 μL, SAP Enzyme (1U / μL) 0.2 μL, ddH2O 1.02 μL. After the reaction system is prepared, seal it tightly with PCR parafilm to prevent sample evaporation, shake and mix, and centrifuge; place the sealed 384-well plate on the ABI9700 PCR instrument for reaction, reaction conditions: 37°C for 40 minutes, 85°C for 5 minutes, and keep at 4°C; The PCR product after treatment with alkaline phosphorylase was obtained and centrifuged for future use.

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Abstract

The invention provides a primer set for amplifying genes related to exercise ability and a detection kit, and relates to the technical field of application of SNPs. The primer set comprises a specificprimer and a single base extension primer; the nucleotide sequence of the specific primer is shown in SEQ ID NO. 1 to 52; and the nucleotide sequence of the single base extension primer is shown in SEQ ID NO.53 to 78 n. The amplification primer and extension primer designed by the invention can detect 26 SNP sites in the same reaction, and are used for developing exercise ability detection kit and guiding reasonable movement. In terms of exercise guidance, the genetic test of the invention can understand the endurance, explosive power, strength and the like of the human body, and provide scientific reference for rational exercise. The movement-related genes are jointly detected and all the items are detected at a time, so that the primer set and the detection kit are most comprehensive incoverage of detection genes in the exercise ability gene detection products.

Description

technical field [0001] The invention belongs to the technical field of SNP application, and specifically relates to a primer set and a detection kit for amplifying genes related to exercise ability. Background technique [0002] Athletic ability refers to the ability of people to engage in various physical activities, including basic abilities such as walking, running, jumping, throwing, climbing, climbing, rolling, rolling, etc., which are indispensable for sustaining life activities, and special athletic abilities for participating in physical training or competitions. , It is the comprehensive expression of human body shape, quality, function, skill and psychological ability. Some studies have pointed out that the former accounts for 2 / 3 and the latter 1 / 3 of the role of genetics and acquired training in becoming an excellent athlete. In recent years, molecular genetics studies have repeatedly confirmed that the occurrence and development of muscle strength, endurance, b...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11C12Q1/686
CPCC12Q1/686C12Q1/6888C12Q2600/156C12Q2600/16C12Q2537/143C12Q2565/627
Inventor 刘文丽武会娟田彦捷洪甜张志超梁丹丹傅莹茜怀雪飞
Owner BEIJING BIOMASION TECH
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