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A kit and application for detecting SNP sites of genes related to calcium demand

A kit and gene technology, applied in the field of kits for detecting SNP sites of genes related to calcium demand, can solve the problems of reduced detection rate and detection efficiency, low specificity and sensitivity, cumbersome operation steps, etc., and achieve a high detection rate High, specific and reproducible results

Active Publication Date: 2021-12-24
HAINAN ZHUJIAN CELL MOLECULE GENETICS INSPECTION CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for the detection of calcium demand-related genes, the existing Sanger sequencing, Taqman method, high-resolution melting curve and other detection methods often have low throughput and cumbersome operation steps. In order to guide calcium supplementation more comprehensively and reasonably, as When the number of detection sites increases, it is prone to low specificity and sensitivity, high sample loading, low detection rate and detection efficiency, and low accuracy for large-throughput detection of multi-gene sites. Effectively used to assess individual differences in calcium requirements to guide genetic testing for calcium supplementation

Method used

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  • A kit and application for detecting SNP sites of genes related to calcium demand
  • A kit and application for detecting SNP sites of genes related to calcium demand
  • A kit and application for detecting SNP sites of genes related to calcium demand

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 - Design of Primer Compositions

[0037] The present invention is based on the optimal selection of 5 SNP locus systems screened out from GC gene, IL-6 gene, PTH gene and VDR gene, and combined with the characteristics of the SNP locus system, a PCR primer composition is designed for amplifying different genes The DNA fragment containing 5 SNP sites, and the combination design of the forward extension primer composition next to the SNP site; at the same time, the extension primers of different lengths are adjusted for different SNP sites, and the five single-base extensions are adjusted The number of poly T modified on the primer and the concentration of the single-base extension primer make the extension products produced by each gene have different molecular weights, so that all primer sequences can accurately and quickly type the sample gene, improving more The accuracy of typing a large number of SNP sites in a reaction system, efficiently and intuitively ...

Embodiment 2

[0041] Preparation and use of embodiment 2-kit

[0042] A. the components and content of the test kit of the present invention include:

[0043] (1) Multiplex PCR amplification reagents: dNTPs, TaqDNA polymerase, PCR reaction buffer, 1x GC-I buffer, Mg2+:

[0044] (2) PCR product purification reagents: AP enzyme, Exol1 enzyme, Seq buffer;

[0045] (3) Multiple single base extension reaction reagent: SNaPshot Multiplex Kit (ABI);

[0046] (4) Extension product purification reagent: SAP enzyme;

[0047] (5) Sequencing denaturation reagents: 120 Liz SIZE STANDARD internal standard, Hi-Di formamide.

[0048] b. Use of kits

[0049] 1. Sample sampling

[0050] Take a saliva sample from the oral test, and carry out DNA extraction to obtain a DNA sample of 1 μl 1% agarose electrophoresis for quality inspection and concentration estimation of the sample, and then divide the sample into 4 equal parts according to the estimated concentration, and dilute them to a concentration of 1...

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Abstract

The present invention provides a kit and application for detecting SNP sites of genes related to calcium demand. The kit includes GC gene RS2282679 sites, IL-6 gene RS1800795 sites, and PTH gene RS6254 sites for detecting calcium demand-related genes. The primer composition of point, VDR gene RS1544410 site and VDR gene RS2228570 site, including 5 pairs of PCR primers and 5 forward extension primer compositions, as shown in SEQ ID No: 1‑15, the present invention can not only quickly synchronize Detect multiple SNP sites, effectively genotype the SNP sites of genes related to calcium demand, and effectively improve the accuracy of typing a large number of SNP sites in one reaction system with a low detection limit, achieving Quickly and accurately give guidance on calcium supplementation.

Description

technical field [0001] The invention relates to the technical field of gene polymorphism detection, in particular to a kit and application for detecting SNP sites of genes related to calcium demand. Background technique [0002] Calcium is the most abundant mineral element in the human body, widely present in all parts of the body. The vast majority of calcium is deposited in the bones and teeth, which not only constitute the scaffold of the human body, but also the storehouse of calcium in the body, while only 1% of calcium exists in the soft tissue, extracellular fluid and blood of the human body. Calcium in soft tissue and blood is also called "mixed calcium pool", which maintains a dynamic balance with calcium in bones and completes a variety of physiological functions. Calcium plays a role in the transmission of nerve impulses, nerve and muscle stress, blood coagulation, and maintenance of heartbeat rhythm. , the activation of human metabolic enzymes and many other phy...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12Q1/6869
CPCC12Q1/6883C12Q1/6869C12Q2600/156C12Q2600/106C12Q2531/113C12Q2537/143C12Q2533/101
Inventor 苏震陈春宝张子询
Owner HAINAN ZHUJIAN CELL MOLECULE GENETICS INSPECTION CENT
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