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Tumor-related SNP site primer composition and application

A primer composition and tumor-related technology, applied in the field of primer composition for tumor-related SNP sites, can solve the problems of less research on tumor SNP sites, and achieve the goal of eliminating non-specific amplification, improving therapeutic effect, and improving accuracy Effect

Inactive Publication Date: 2021-09-21
BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few studies on tumor-associated SNP sites in the prior art

Method used

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  • Tumor-related SNP site primer composition and application
  • Tumor-related SNP site primer composition and application
  • Tumor-related SNP site primer composition and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Primer Composition

[0064] The primer composition described in this application is specifically shown in Table 2 below.

[0065] Table 2 Primer Composition

[0066]

[0067]

[0068]

[0069]

[0070]

[0071]

[0072]

[0073]

Embodiment 2

[0074] Embodiment 2 library construction and detection

[0075] 1. Genomic DNA Extraction

[0076] Genomic DNA (gDNA) was extracted from saliva or blood according to the operating steps of the commercial genomic DNA extraction kit, and quantified using Qubit (Life Technologies).

[0077] 2. The first round of multiplex PCR reaction

[0078] 1.1. The reaction system is shown in Table 3 below.

[0079] Table 3 reaction system

[0080] Reagent Volume(μL) wxya 2 o

4-x Enhancer NB(1N) 5 Primer (Table 2) 2.4 gDNA X (30ng) IGT-EM101 polymerase mixture 3.6

[0081] Wherein, the final concentration of each primer in the Primer primer mixture is 0.5 pmol.

[0082] 1.2. The reaction program is: 95°C for 3min30s; 98°C for 20s, 60°C for 8min, 18cycle; 72°C for 5min.

[0083] 3. The first round of magnetic bead purification

[0084] 3.1. Add 15 μL of AMPure XP magnetic beads equilibrated at room temperature to 15 μL of PCR product, an...

Embodiment 3

[0119] The corresponding relationship between tumors, genes and detection sites is shown in Table 5 below.

[0120] table 5

[0121]

[0122]

[0123]

[0124]

[0125]

[0126] Experimental Example 1 Accuracy Detection

[0127] The detection method described in this application was used to detect the above 168 sites, and Sanger sequencing was used for verification at the same time, the results are shown in Table 6 below.

[0128] Table 6 Accuracy test results

[0129]

[0130]

[0131]

[0132]

[0133]

[0134] It can be seen from Table 6 that the accuracy of the primer composition and detection method described in this application is 100%.

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Abstract

The invention provides a tumor-related SNP site primer composition and application thereof, and belongs to the technical field of molecular biology. According to the invention, a group of SNP site amplification primers are designed and developed through an NGS method, and through a bioinformatics analysis method, the change of a gene structure related to tumors in an individual body is detected, the disease risk of the individual body is predicted, and the prevention and / or treatment effect of the tumors is improved. The evaluation system and the detection method disclosed by the invention are low in cost, simple and convenient to operate, good in sensitivity, high in accuracy and good in repeatability, and have great clinical application value.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a primer composition and application of a tumor-associated SNP site. Background technique [0002] Tumor is an abnormal lesion formed by the abnormal proliferation of cells due to abnormal gene expression regulation of local tissue cells under the action of carcinogenic factors, which can not normally regulate cell growth. It can be divided into benign tumors and malignant tumors. The former generally does not Recurrence, non-metastasis, only blockage and extrusion on tissues and organs, which can be removed by surgical treatment; the latter is also called cancer (Cancer), cancer cells not only grow out of control, but also invade the surrounding area unlimitedly and endlessly Normal tissues are transferred to other tissues of the body through the internal circulatory system. The invasion of cancer cells leads to pain, bleeding, necrosis, infection, emaciation, weakness...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6858C12N15/11
CPCC12Q1/6886C12Q1/6858C12Q2600/156C12Q2531/113C12Q2537/143C12Q2537/165C12Q2535/122C12Q2563/143C12Q2563/149
Inventor 方向东渠鸿竹单广乐刘彦霞王晨
Owner BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
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