Method for extracting genomic DNAs from preserved animal samples

A genome and DNA damage technology, applied in the field of molecular biology, can solve the problems of improving the yield of nucleic acid extraction, cumbersome process of dewaxing, etc., achieving low extraction cost, short operation time, eliminating C>T and G>A artificial effect of mutation

Inactive Publication Date: 2019-04-09
BEIJING USCI MEDICAL LAB CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The dewaxing method currently used in mature kits mainly uses organic solvents such as xylene that have a pungent taste and are more harmful to the human body for dewaxing, and the dewaxing process is cumbersome, and there are many types of reagents used. Takes at least 20 minutes
At the same time, since formaldehyde-fixed samples will randomly produce C-T conversion, and there is loss of nucleic acid in animal samples during preservation, and the extraction methods in the prior art have no effect on the generation of such artificial mutations, DNA damage and formaldehyde fixation or Therefore, it is urgent to develop a method for extracting genomic DNA that can obtain high yield, high quality, and provide accurate DNA base information.

Method used

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  • Method for extracting genomic DNAs from preserved animal samples
  • Method for extracting genomic DNAs from preserved animal samples

Examples

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Embodiment 1

[0057] This embodiment provides a method for extracting genomic DNA from paraffin-embedded animal tissue samples. The extracted samples are the surgical tissue paraffin patch derived from breast tissue of breast cancer patients and the puncture tissue paraffin wax derived from left lung lobe tissue of lung cancer patients. patch, the specific method is as follows:

[0058] (1) Use a clean scalpel blade to scrape a tissue paraffin section sample with an area >10mm×10mm and a thickness of about 5-10μm, transfer it to a prepared 1.5mL centrifuge tube, centrifuge at 12000rpm for 3min, and centrifuge the scraped tissue into the tube. end;

[0059] (2) Add 160 μL of dewaxing solution (the dewaxing solution is n-hexadecane) to the above-mentioned centrifuge tube containing paraffin-embedded tissue. If the sample volume is large or the paraffin content is large, double the amount of dewaxing solution can be used for processing ;

[0060] (3) Add proteinase K to a final concentration...

Embodiment 2

[0072] This embodiment provides a method for extracting genomic DNA from formaldehyde-soaked animal tissues. The difference from the method provided in Example 1 is only step (1) and step (2). In this embodiment, the steps of embodiment (1) (1) and step (2) are combined into step (1): Scrape the formaldehyde-soaked surgical tissue block with a clean scalpel blade, transfer to a prepared 1.5ml centrifuge tube, centrifuge at 12000rpm for 3min, and centrifuge the scraped tissue to tube bottom. All the other steps are the same as in Example 1. The sample extracted in this embodiment is formaldehyde-soaked surgical tissue derived from breast tissue of a breast cancer patient.

Embodiment 3

[0074] This embodiment provides a method for extracting genomic DNA of animal pleural effusion, the difference with the method provided in Example 1 is only step (1) and step (2), in the present embodiment, the step of embodiment (1) ( 1) and step (2) are combined into step (1): Take 200 μl of pleural and ascites fluid into a prepared 1.5ml centrifuge tube, centrifuge at 12000 rpm for 3 minutes, and centrifuge the scraped tissue to the bottom of the tube. All the other steps are the same as in Example 1. The extracted samples in this embodiment are pleural and ascites from lung cancer patients.

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Abstract

The invention relates to the technical field of molecular biology, and in particular to a method for extracting genomic DNAs from preserved animal samples. The method for extracting genomic DNAs frompreserved animal samples comprises a step of adding repair solution to a lysed sample for mutation base elimination and DNA damage repair; and the repair solution comprises a UNG enzyme. The genomic DNA extraction method provided by the invention is capable of extracting high-yield and high-quality genomic DNAs from limited preserved animal samples and effectively eliminating C)T and G)A human mutations, and has the advantages of being short in operation time, simple and convenient to operate, low in extraction cost, high in unit throughput of extraction process and high in safety of operation.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a method for extracting genomic DNA from preserved animal samples. Background technique [0002] In the process of scientific research and clinical practice, animal samples such as animal tissues obtained cannot be tested immediately, but animal samples need to be properly processed for short-term or long-term storage. At present, the preservation of animal tissues mainly adopts formalin immersion or preparation of animal tissues as formalin-fixed and paraffin-embedded (Formalin-Fixed and Parrffin-Embedded, FFPE) samples. Formalin can irreversibly cross-link with the amino groups of protein amino acids, and is superior to other fixatives in terms of maintaining the integrity of tissue cell structure, antigen measurability, and tissue permeability. Formalin-fixed or formalin-paraffin-embedded physiological or pathological samples contain valuable and widely sourced mate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1006C12N15/1013C12N15/1024
Inventor 孟岳峰孙晴晴潘晓西王建伟伍启熹刘倩刘珂弟唐宇
Owner BEIJING USCI MEDICAL LAB CO LTD
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