Method for extracting desoxyribonucleic acid from formalin fixed and paraffin embedded tissues

A deoxyribonucleic acid and paraffin-embedded technology, applied in the field of nucleic acid applications in biology, can solve the problems of unsuitable handling of a large number of specimens, injury to the operator's body, and high cost of proteinase K. Simple to use effects

Active Publication Date: 2011-08-10
TIANGEN BIOTECH BEIJING
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Problems solved by technology

[0003] At present, the traditional paraffin-embedded tissue DNA extraction mostly adopts the method of xylene dewaxing and proteinase K incubation. The whole extraction process of this method is long, and proteinase K is expensive, and the operation is very cumbersome. Time-consuming and reagents are required, an...

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  • Method for extracting desoxyribonucleic acid from formalin fixed and paraffin embedded tissues
  • Method for extracting desoxyribonucleic acid from formalin fixed and paraffin embedded tissues
  • Method for extracting desoxyribonucleic acid from formalin fixed and paraffin embedded tissues

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[0031] Example: DNA extraction from formalin-fixed paraffin-embedded tissue sections of mouse liver.

[0032] 1. Extraction of DNA from formalin-fixed paraffin-embedded tissues

[0033] (1) Take 2 to 4 paraffin-embedded tissue sections with a thickness of 10 microns (formalin immersion time is 8 hours and 24 hours respectively), and put them in 500 microliters of sodium hydroxide solution with a molar concentration of 0.1 , then add 50 microliters of sodium lauryl sulfate with a mass percentage concentration of 20% and 50 microliters of dithiothreitol with a molar concentration of 0.001, mix well, and incubate at 100°C for 30 minutes to obtain a lysate ;

[0034] (2) Carry out centrifugal stratification to above-mentioned lysate, the rotating speed of centrifugal stratification is 1,3200 rev / mins, and centrifugal stratification time is 5 minutes, takes out the middle layer after centrifugal stratification, adds 500 microliters of absolute ethanol, Thoroughly oscillate and mi...

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Abstract

The invention relates to a method for extracting desoxyribonucleic acid from formalin fixed and paraffin embedded tissues, and belongs to the technical field of nucleic acid application in biology. The method comprises the following steps of: preparing special lysis solution, adding the lysis solution into paraffin section tissues, boiling at high temperature for 30 minutes, and centrifuging to take supernate; adding absolute ethanol for uniform mixing, and adding the mixed solution into a silicon membrane absorption column for centrifuging; rinsing by using protein-free liquid and salt-free rinsing liquid; and eluting by using elution buffer. In the method, a toxic reagent of dimethylbenzene is not used for dewaxing, and harm to bodies of experimenters is avoided; and precious protease Kis not needed to perform long-time incubation enzymolysis, the operation is simple and quick, the extracted desoxyribonucleic acid in genome is high in quality and stability, and the cost and time can be saved to the greatest degree. The extracted product is subjected to polymerase chain reaction (PCR) detection, long segments with about 750pb can be obtained through amplification, and the work in the aspects such as scientific research, biomedicine and the like is greatly facilitated.

Description

technical field [0001] The invention relates to a method for extracting deoxyribonucleic acid from formalin-fixed paraffin-embedded tissues, belonging to the technical field of nucleic acid applications in organisms. Background technique [0002] Formalin is not only a preservative, but also has a good effect on maintaining the shape of many biological specimens. It is superior to other fixatives in terms of maintaining the integrity of tissue cell structure, antigen measurability, and tissue permeability. Therefore, it has been widely used in the research of biological evolution, phylogeny, endangered species protection, population genetics and other disciplines, as well as in clinical testing and medical science research. Therefore, a relatively simple, effective, safe and feasible method for extracting high-quality genomic deoxyribose nucleic acid (Deoxyribonucleic acid, hereinafter referred to as DNA) from formalin fixed and paraffin embedded tissues (FFPE, hereinafter ...

Claims

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Application Information

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IPC IPC(8): C07H21/04C07H1/08
Inventor 韩典霖俞萍李晓晨孙克非
Owner TIANGEN BIOTECH BEIJING
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