Method for extracting mammary tissue RNA

An extraction method and breast tissue technology, applied in the field of RNA extraction and purification, can solve the problems of unextracted RNA, low quality, low concentration, etc., and achieve the effect of reducing the amount of tissue used, less tissue, and ensuring the concentration

Pending Publication Date: 2019-09-24
WEST CHINA HOSPITAL SICHUAN UNIV
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Problems solved by technology

[0004] Normal mammary gland tissue for RNA extraction is scarce, and people often get low-quality and low-concentration RNA using conventional RNA extraction methods, and sometimes even no RNA can be extracted

Method used

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  • Method for extracting mammary tissue RNA

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Embodiment 1

[0024] Embodiment 1 breast tissue RNA extraction method of the present invention

[0025] 1 Pretreatment of tissue lysis: section identification and section sampling by frozen microtome

[0026] 1) Take out the breast tissue (including breast glandular tissue, adipose tissue and connective tissue) samples from the -80 degree refrigerator;

[0027] 2) Embed the sample in a frozen microtome to make frozen sections;

[0028] 3) Section identification: HE staining determines the area where the mammary gland cells are located in the tissue, and cuts out the section area with mammary gland tissue.

[0029] 4) Weighing to determine that the amount of tissue used for a single extraction is not greater than 30 mg.

[0030] 2 Tissue sample lysis and RNA extraction

[0031] Put the aforementioned sliced ​​tissue material into the cell lysate for RNA extraction (the lysate in the QIAGAN RNeasy Mini Kit), and blow it off with a pipette.

[0032] Use the RNA extraction kit (QIAGAN RNeas...

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Abstract

The invention provides a pretreatment method for extracting mammary tissue RNA. The method comprises the steps that a mammary tissue frozen section material is prepared first, mammary gland tissue in the section material is indentified through HE staining, and the mammary gland tissue is cut off. The mammary gland tissue is directly used for cell lysis and RNA extraction, and then the high-quality and high-concentration RNA can be obtained. By means of the method, the problem can be solved that the mammary tissue for extracting the RNA is scarce to cause low RNA extracting quality.

Description

technical field [0001] The invention relates to the field of RNA extraction and purification. Background technique [0002] With the development of modern medicine, biomarkers have gradually become important reference indicators for disease diagnosis and treatment. [0003] In the detection of breast cancer, it is necessary to extract and detect RNA in cancerous and normal breast glandular tissues (rather than adipose and connective tissues). Cancerous breast glandular tissue is relatively easy to obtain, but normal breast glandular tissue is scarce. The reason is: breast cancer patients who need breast-conserving surgery have less normal breast tissue, and it is impossible to sample a large number of them; and cases of total mammectomy and neoadjuvant chemotherapy are rare. [0004] Normal mammary gland tissue for RNA extraction is scarce, and people often obtain low-quality and low-concentration RNA using conventional RNA extraction methods, and sometimes even no RNA can...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 陈洁王竹贺涛王艳萍王宇范园胡靖睿
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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