Method for extracting DNA from formalin-fixed tissue

A formalin and tissue technology, applied in the field of DNA extraction from tissue, can solve the problems of DNA separation failure, inhibition of PCR reaction, expensive Triton-100, etc.

Inactive Publication Date: 2010-06-23
HUAZHONG UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

In addition, formalin may also have an inhibitory effect on the digestive juice and affect the lysis of cells, which may also lead to the failure of DNA isolation
In 2003, Jia Yi et al. reported using Triton-100 to extract DNA from formalin-fixed tissue, but Triton-100 is expensive and has an inhibitory effect on PCR reactions, so it needs to be improved

Method used

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  • Method for extracting DNA from formalin-fixed tissue
  • Method for extracting DNA from formalin-fixed tissue
  • Method for extracting DNA from formalin-fixed tissue

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Embodiment 1

[0023] The method for extracting DNA from formalin-fixed tissue provided by the invention comprises the following steps:

[0024] 1. Cut out about 30mg of fixed tissue and put it into a 1.5ml EP centrifuge tube;

[0025] 2. Shred as much as possible so that the tissue cells can be completely lysed and digested in step 6;

[0026] 3. Rinse the tissue fragments with graded ethanol (30%, 40%, 50%, 60%, 70%, 80%, 90% and 100%) for 20 minutes each time, and centrifuge at 3000g for 10 minutes to gradually remove Fu Marlin;

[0027] 4. Put it in a silica gel desiccator and let it stand overnight to completely remove formalin;

[0028] 5. Add 300 μl of cell lysate (500 mmol / L Tris pH8.0, 20 mmol / L EDTA, 10 mmol / L NaCl, 1% SDS) and 20 μl of proteinase K (20 mg / ml);

[0029] 6. Shake and mix for 1 minute, then place at 55°C for 2 days to digest the protein, and shake 2-3 times during the process to promote digestion;

[0030] 7. Take out the sample, mix gently after cooling down to ...

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Abstract

The invention provides a method for extracting DNA from formalin-fixed tissue. The method combines a formalin removal method with a conventional DNA isolation method, can extract high-quality genomic DNA from the formalin-fixed tissue, and has the characteristics that: the method has low cost and is easy to popularize and apply; and the obtained DNA is applicable for PCR amplification, and the like.

Description

technical field [0001] The invention belongs to biotechnology and relates to a DNA extraction method, in particular to a method for extracting DNA from tissues Background technique [0002] Formalin-fixed tissue is a precious genetic resource, and the genetic material DNA in them can be well preserved for a long time. We can separate and purify it and conduct genetic testing on it, which undoubtedly expands our research materials. The scope also provides conditions for us to conduct retrospective research. However, isolating DNA from these tissues does not copy the method of isolating DNA from fresh tissue. Formalin fixation makes DNA isolation difficult. Formaldehyde, the main component of formalin fixative, can cause extensive cross-linking between DNA and protein to form a firm complex, which can hinder the digestion of tissue by proteinase K, thereby affecting DNA extraction. In addition, formalin may also have an inhibitory effect on the digestive juice and affect th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H21/04C12N15/10
Inventor 陈会平杨真荣
Owner HUAZHONG UNIV OF SCI & TECH
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