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High-flux screening method of hereditary breast cancer mutant gene

A technology for mutating genes and breast cancer, which is applied in biochemical equipment and methods, and microbial measurement/inspection, etc., can solve problems such as PTEN enzyme inactivation, and achieve strong predictability and high accuracy

Pending Publication Date: 2018-02-23
JIAXING YUNYING MEDICAL INSPECTION CO LTD
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  • Abstract
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AI Technical Summary

Problems solved by technology

The mutation of PTEN will cause the inactivation of PTEN enzyme, activate the PI3K-AKT signaling pathway and cause cancer

Method used

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  • High-flux screening method of hereditary breast cancer mutant gene
  • High-flux screening method of hereditary breast cancer mutant gene

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Experimental program
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Effect test

Embodiment Construction

[0043] A kind of screening method of hereditary breast cancer mutation gene of the present invention, comprises the steps:

[0044] 1. Sample library preparation:

[0045] (1) Ultrasound fragmentation: the initial amount is 3ug, add nuclease-free water to 100ul and dilute to 30ng / ul. SCIENTZ08-Ⅲ cup-type ultrasonic cell pulverizer was used for ultrasonic fragmentation, and the setting parameters were: power 70%, interrupt for 3s, stop for 1s, and cycle for 30-60min.

[0046] (2) Take out the magnetic beads half an hour in advance to return to room temperature, shake and mix, take 180ul of magnetic beads and add them to the PCR product interrupted by ultrasound, beat and mix, and incubate in the greenhouse for 5 minutes.

[0047] (3) Put the PCR single tube on the magnetic stand, let it stand for 5 minutes, remove the supernatant, keep the PCR tube on the magnetic stand, add 200ul of 80% ethanol (newly prepared) to rinse, let stand for 30s, remove the supernatant, 80 Rinse tw...

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Abstract

The invention discloses a rapid and accurate screening method of a hereditary breast cancer mutant gene capable of covering all exome regions of the latest hereditary breast cancer. The method disclosed by the invention is characterized in that 3 to 5 ml of blood can be extracted from the individual human body, 3 to 5 micro g gDNA is extracted from the blood and ultrasonically broken and amplifiedso as to establish a human whole-genome library, then a hereditary breast cancer mutant gene scanning kit of the invention is used for capturing relevant disease genes, then a new-generation sequencing instrument is used for performing the high-flux sequencing to analyze and find out the mutation information related to the genes, thereby achieving a purpose of screening the hereditary breast cancer mutant genes.

Description

technical field [0001] The invention relates to a gene screening method, in particular to a screening method for hereditary breast cancer mutation gene. technical background [0002] Breast cancer is a common malignant tumor in women. It occurs in the glandular epithelial tissue of the breast. The incidence rate accounts for 7-10% of all kinds of malignant tumors in the whole body. It seriously affects the physical and mental health of women. In my country, the incidence of breast cancer is growing the fastest, increasing at a rate of 3-4% every year, accounting for the first place in women's cancer deaths. Breast cancer cells are loosely connected and easy to fall off. Once the cancer cells fall off, they will spread throughout the body with blood or lymph fluid, forming metastasis, which is life-threatening. The natural course of breast cancer is usually very long, and the average doubling time of cancer cells is 90 days. People often find that they have lymph node meta...

Claims

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Application Information

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IPC IPC(8): C12Q1/6869
CPCC12Q1/6869C12Q2531/113C12Q2535/122
Inventor 张道允巩子英叶建伟王伟
Owner JIAXING YUNYING MEDICAL INSPECTION CO LTD
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