Method for rapidly and efficiently extracting genomic DNA of mammal ear tissue

A mammalian and genome technology, applied in the field of molecular biology, can solve the problems of long time-consuming, low concentration of DNA extraction, etc., and achieve the effect of improving the yield

Inactive Publication Date: 2016-06-08
GANSU AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The technical problem to be solved by the present invention is to provide a method for quickly and efficiently extracting genomic DNA f...

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  • Method for rapidly and efficiently extracting genomic DNA of mammal ear tissue
  • Method for rapidly and efficiently extracting genomic DNA of mammal ear tissue
  • Method for rapidly and efficiently extracting genomic DNA of mammal ear tissue

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Embodiment Construction

[0020] The present invention and its effects will be further described below.

[0021] A method for quickly and efficiently extracting mammalian ear tissue genomic DNA, the specific steps of the method are as follows:

[0022] 1) Cut the tissue sample into pieces in a sterilized centrifuge tube and quickly add pre-cooled TE to fully break up the cells and protect the DNA in the solution;

[0023] 2) The digestion of tissue lysate eliminates the overnight digestion with proteinase K;

[0024] 3) Tris-saturated phenol extraction;

[0025] 4) Mixed extraction of Tris-saturated phenol and chloroform isoamyl alcohol;

[0026] 5) Chloroform isoamyl alcohol extraction;

[0027] 6) Precipitate DNA with sodium acetate (NaAc) and -20°C pre-cooled absolute ethanol;

[0028] 7) Ethanol rinse;

[0029] 8) TE dissolves the DNA and saves it.

[0030] The specific implementation is as follows:

[0031] 1) Preparation of TE lysate (pH8.0): 1MTris-HCl (pH8.0) 5mL, 0.5MEDTA (pH8.0) 2mL, a...

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Abstract

The invention discloses a method for rapidly and efficiently extracting genomic DNA of mammal ear tissue. The method comprises steps as follows: 1) tissue disruption and DNA protection; 2) tissue lysate digestion; 3) Tris-saturated phenol extraction; 4) Tris-saturated phenol and chloroform/isoamyl alcohol mixed extraction; 5) chloroform/isoamyl alcohol extraction; 6) precipitation of DNA with NaAc and fully precooled (subzero 20 DEG C) absolute ethanol; 7) ethanol rinsing; 8) dissolution of DNA with a TE buffer solution and preservation. The yield of DNA is significantly increased (the concentration of DNA is 112-851 ng/mu L), meanwhile, the time spent on addition of proteinase K for digestion overnight (longer than or equal to 12 h) after cell disruption during DNA extraction is saved, and about 3 h is spent from tissue disruption to genomic DNA extraction.

Description

technical field [0001] The invention relates to a method for rapidly and efficiently extracting mammalian ear tissue genomic DNA, belonging to the technical field of molecular biology. Background technique [0002] Genomic DNA extraction is an important technical link in molecular biology research, and is widely used in germplasm resource evaluation and breeding research. In order to facilitate the search for animal blood relationship and management records, ear clipping is often used after the animal is born. The method is used to number each individual. At this time, the collection of ear defect tissue can meet the research of large sample size without harming the health of animals. The current method of extracting genomic DNA from mammalian tissues usually adopts the traditional phenol-chloroform extraction method and the kit method. The former is directly digested with proteinase K after the tissue is broken, which takes a long time (≥12h) and the cell is not fully broke...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 滚双宝马艳萍杨巧丽
Owner GANSU AGRI UNIV
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