Primer set, application, product and method for detecting SNP sites related to nutrient metabolism ability

A technology of metabolism and nutrient metabolism, applied in the field of primer sets for detecting SNP sites related to nutrient metabolism, can solve the problems of high sample loading, low throughput, complicated operation, etc., to shorten the detection cycle, reduce the detection cost, Accurate effect

Active Publication Date: 2020-07-24
NANJING SIMCERE MEDICAL LAB CO LTD +2
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above methods all have some shortcomings or deficiencies. For example, although Sanger sequencing is the gold standard for sequencing and can detect unknown mutations, this method has disadvantages such as low throughput, cumbersome operation, and high sample loading; Taqman method can only detect Known mutations, and the price of probe synthesis is high; ARMS method has

Method used

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  • Primer set, application, product and method for detecting SNP sites related to nutrient metabolism ability
  • Primer set, application, product and method for detecting SNP sites related to nutrient metabolism ability
  • Primer set, application, product and method for detecting SNP sites related to nutrient metabolism ability

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Example 1 Design and optimization of primers, establishment of reaction system

[0077] 1.1 Design and optimization of primers

[0078] Through the primer design software (Assay Design Suite) on the MassARRAY website, adjust the relevant parameters, complete the 51-site PCR and UEP primer design, export the designed primers and various parameter files, and synthesize the primers. Prepare amplification primer MIX and extension primer MIX according to the primer configuration table, and fine-tune the extension primer MIX until it meets the requirements. Then proceed to primer testing and optimization. Specific steps are as follows:

[0079] (1) Dilute the genomic DNA sample to 10ng / μl, and prepare the PCR reaction MIX according to the table below (the following is a single sample volume)

[0080] Table 3 PCR reaction system

[0081]

[0082] Seal the film, vortex and mix for 30 seconds, centrifuge at 4000 rpm for 1 minute, put the plate on the PCR machine for the following therm...

Embodiment 2

[0117] Example 2 Verification of the reaction system

[0118] In the system optimization process of the present invention, each detection site has 2 to 3 samples verified by Sanger sequencing, and the comparison results are consistent, confirming that the detection result of the present invention is accurate. The present invention performs a series of verification experiments after confirming the optimal reaction system, including accuracy, precision, and personnel comparison and comparison experiments between different batches of primers. The specific verification scheme is as follows:

[0119] (1) Accuracy experimental verification scheme: One sample from each of 51 sites is selected for Sanger sequencing, and the results of Sanger sequencing and MassARRAY are compared. If the consistency is greater than 95%, the verification is passed.

[0120] (2) Accuracy experimental verification scheme: Pick 3 exceptions of peripheral blood samples and 3 corresponding buccal swab samples, eac...

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Abstract

The invention relates to the technical field of biology and provides a nutrition metabolic capacity related SNP site detection primer group, application, product and method. The nutrition metabolic capacity related SNP site detection primer group can be used for detecting vitamin A metabolic capacity, vitamin B6 metabolic capacity, vitamin B12 metabolic capacity, vitamin C metabolic capacity, vitamin D metabolic capacity, vitamin E metabolic capacity, elemental calcium metabolic capacity, elemental iron metabolic capacity, elemental zinc metabolic capacity, elemental selenium metabolic capacity, elemental iodine metabolic capacity, elemental sodium metabolic capacity, alcohol metabolic capacity, folic acid metabolic capacity, lactose metabolic capacity, unsaturated fatty acid metabolic capacity, saturated fatty acid metabolic capacity, cholesterol metabolic capacity, carbohydrate metabolic capacity and caffeine metabolic capacity. By the primer group, specific detection can be realized, high accuracy is achieved, a detection period can be shortened sharply, and detection cost is reduced.

Description

Technical field [0001] The present invention relates to the field of biotechnology, in particular to a primer set, application, product and method for detecting SNP sites related to nutrient metabolism. Background technique [0002] Nutrient elements play an irreplaceable role in the human body and are necessary to maintain good health. They mainly include vitamins, trace mineral elements and other nutrients such as lactose, cholesterol, and carbohydrates. Lack of nutrients and excessive supplementation can cause physiological disorders or cause other diseases. A large number of studies have shown that the absorption and metabolism of a variety of nutrient elements are related to gene polymorphism, and gene mutations will cause differences in the activity of synthetic metabolic enzymes, which will cause differences in the body's metabolism and absorption of nutrients. Therefore, genetic testing can assess the individual's ability to absorb and metabolize different nutrients, and...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 李小方陈珊珊赵妍李翔李杜衡肖念清任用
Owner NANJING SIMCERE MEDICAL LAB CO LTD
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