Changbai mount rhododendron chrysanthum pall genome DNA extraction method

A genome and rhododendron technology, applied in the field of Changbai Mountain cowhide rhododendron genome DNA extraction, can solve the problems of easy degradation, difficult genome extraction, low purity, etc., and achieve high extraction efficiency, simple and feasible operation, and high purity.

Inactive Publication Date: 2013-05-01
JILIN NORMAL UNIV
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  • Abstract
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  • Application Information

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Problems solved by technology

[0005] In view of the above-mentioned deficiencies, the object of the present invention is to overcome the shortcomings of the prior art in the Changbai Mountain rhododendron genomic DNA extraction method, genome extractio

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  • Changbai mount rhododendron chrysanthum pall genome DNA extraction method
  • Changbai mount rhododendron chrysanthum pall genome DNA extraction method

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Embodiment Construction

[0026] Below in conjunction with implementation case the present invention will be further described:

[0027] 1. Genomic DNA of Changbai Mountain Rhododendron extracted by CTAB-SDS method

[0028] (1) Improving the CTAB method to extract plant genomic DNA

[0029] 1. Take about 0.4 g of fresh leaves of Rhododendron chinensis in Changbai Mountain, grind them into powder with liquid nitrogen, put them in a 2 ml centrifuge tube, add 700 ul of o C preheated CTAB extraction buffer (2% CTAB, 100 mmol L -1 Tris-HCl pH 8.0, 20mmol L -1 EDTA; 1.4 mol L -1 NaCl, 4% PVP), and 30 ul β-mercaptoethanol was added before trituration.

[0030] 2. After mixing quickly and fully, put it in a water bath at 65 o C incubation for 4 hours, shaking gently every 30 minutes.

[0031] 3. Add an equal volume (730 ul) of phenol: chloroform (1:1) mixture, mix the mixture by inversion, 18 o Centrifuge at 12,000 rpm for 10 min; repeat once.

[0032] 4. Take the supernatant, add 3 times the volu...

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Abstract

The invention belongs to the technical field of genetic engineering, and discloses an alpine plant Changbai mount rhododendron chrysanthum pall genome DNA extraction method. According to the method, a CTAB method and a SDS method are combined at the first time to extract and purify Changbai mountain rhododendron chrysanthum pall genome DNA; in the extraction method, a PVP use amount in a CTAB extraction buffer solution is 4% and is increased by 2% compared to a PVP use amount in the conventional method, a 2 beta-mercaptoethanol use amount is 4.3% and is increased by about 1 time compared with a 2-mercaptoethanol use amount in the conventional method, and a 65 DEG C warm bath time is prolonged to 4 h from 1 h of the conventional method; and the SDS method is combined to purify the extracted genome so as to successfully extract the high purity and good quality Changbai mountain rhododendron chrysanthum pall genome DNA. With the present invention, disadvantages of difficult extraction, low purity of the extracted genome and easy degradation of the existing alpine plant are overcome, and important significances are provided for subsequent molecular level researches and alpine plant genome researches.

Description

technical field [0001] The invention relates to a method for extracting genome DNA of alpine plants, belonging to the technical field of genetic engineering, in particular to a method for extracting genome DNA of Changbai Mountain cowhide rhododendron. Background technique [0002] Due to the harsh environmental conditions of alpine plants, alpine plants are generally considered to be the most extreme creatures on land, but alpine plants also have rich and valuable genetic resources. Since 1896, as a hot and difficult point in the study of biological evolution, the adaptation mechanisms and strategies of alpine and polar plants to habitats have been the focus of researchers. The extraction technology of alpine plant genomic DNA is one of the basic technologies for plant molecular biology research. The yield and quality of DNA directly affect the success or failure of subsequent experiments. Discussing the extraction method of alpine plant genomic DNA is a very meaningful wo...

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Application Information

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IPC IPC(8): C12N15/10
Inventor 徐洪伟周晓馥未晓巍
Owner JILIN NORMAL UNIV
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