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BRCA1/2 genovariation combined detection kit and application thereof

A joint detection and gene variation technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of high price, low sensitivity, and time-consuming, and achieve low cost, simple experimental process, The effect of low demand

Active Publication Date: 2017-08-22
浙江绍兴鼎晶生物医药科技股份有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current methods for detecting BRCA1 / 2 gene mutations mainly include Sanger sequencing, qPCR method or MLPA, but the sequencing throughput of these methods is low, the sensitivity is not high, and it takes a lot of time and is expensive

Method used

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  • BRCA1/2 genovariation combined detection kit and application thereof
  • BRCA1/2 genovariation combined detection kit and application thereof
  • BRCA1/2 genovariation combined detection kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] HORIZON BRCA Somatic Multiplex Reference Standard was used for combined detection of multi-gene driver mutations.

[0091] 1. Dilute the gDNA according to the kit and method of the present invention, and use the qubit dsDNA HS Assay Kit quantitative kit to accurately quantify the diluted gDNA so that the DNA concentration is about 3 ng / uL.

[0092] A. Target DNA amplification;

[0093] According to the following components, prepare a PCR system to amplify the target fragment.

[0094]

[0095]

[0096] Carry out PCR amplification according to the following amplification conditions:

[0097]

[0098] After PCR amplification, the two sets of amplification products of the same template DNA sample were mixed.

[0099] B. Partial digestion of primer sequences:

[0100] Add 2 μL of digestive enzyme to the amplified product, and carry out the digestion reaction under the following conditions. Can be performed in a PCR machine.

[0101] temperature time ...

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Abstract

The invention provides a BRCA1 / 2 genovariation combined detection kit and belongs to the field of genovariation detection. The kit comprises a BRCA1 gene primer group, a BRCA2 gene primer group, polymerase, a PCR reaction liquid, a digestive enzyme, ligase, a ligase buffer solution, an attachment joint, a fluorescence probe, a Raq enzyme, a qPCR primer and a qPCR reaction liquid, wherein the Raq enzyme, the qPCR primer and the qPCR reaction liquid are used in a qPCR reaction. The combined detection kit can detect mutation sites of BRCA1 / 2 genes of breast cancer at the same time, the experimental result is high in sensitivity, the 5% or below of mutation rate can be detected, the detection sensitivity is greatly improved, the problems in the prior art are overcome, and meanwhile, the whole experiment process meets the requirements for mass and quick detection.

Description

technical field [0001] The invention belongs to the technical field of gene variation detection, and in particular relates to a combined detection kit for BRCA1 / 2 gene variation and an application thereof. Background technique [0002] In 1990, researchers discovered a gene closely related to hereditary breast cancer, named breast cancer 1 gene (BRCA1), located on chromosome 17; Another gene associated with breast cancer, called BRCA2, was discovered and the two genes are commonly referred to collectively as BRCA1 / 2. [0003] BRCA is a tumor suppressor gene, which plays an important role in regulating cell replication, DNA damage repair, and normal cell growth; if the BRCA gene is mutated, it will lose the function of inhibiting tumorigenesis. There are hundreds of types of BRCA mutations, which are related to the occurrence of many cancers in the human body, among which the most closely related is breast cancer, followed by ovarian cancer. [0004] Breast cancer is one of...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/156C12Q2600/16
Inventor 朱凯陈悦科陈成刘璐吴燕灵
Owner 浙江绍兴鼎晶生物医药科技股份有限公司
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