Method for extracting DNA from Xinjiang garlic wild relative species

An extraction method and technology of related species, applied in the field of plant molecular biology research, to achieve the effects of low environmental risk, short time consumption, and cost reduction

Inactive Publication Date: 2018-01-05
XINJIANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the technical problems existing in Xinjiang wild relatives of garlic, the purpose of this invention is to provide a simple and practical method for extracting genomic DNA from wild relatives of garlic seeds

Method used

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  • Method for extracting DNA from Xinjiang garlic wild relative species
  • Method for extracting DNA from Xinjiang garlic wild relative species
  • Method for extracting DNA from Xinjiang garlic wild relative species

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 A method for extracting genomic DNA from seeds of wild relatives of garlic in Xinjiang, referring to the method of Wang Zifeng with slight modifications, includes the following steps:

[0037] (1) From early to mid-August, in the original environment of Tianchi Lake in Tianshan Mountains (43°54′2.41″N, 88°07′18″E; altitude 1891m), the seeds of wild relatives of garlic (Garlicum prismatica) were used as materials, and the phenology The stage is in the fruit ripening stage. When the capsules are cracked and the pericarp is withered yellow, collect the mature seeds, put them in a kraft paper bag (envelope), bring them back to the laboratory, and dry them at room temperature for 7 days.

[0038] (2) Determination of seed water content: Randomly select the collected seeds, weigh them with an electronic balance, and record it as W 1 , and then put the seeds in an oven at 120°C, take them out every 4 hours and weigh them after the temperature drops to room temperatur...

Embodiment 2

[0050] The mensuration of embodiment 2 seed water content

[0051] 1. Materials

[0052] Randomly select about 1000 grains of rib-leaved garlic seeds that are uniform in size, plump, and dried naturally, and weighed with an electronic balance, recorded as W 1 , and then put the seeds in an oven at 120°C, take them out every 4 hours and weigh them after the temperature drops to room temperature, and repeat it several times until the weight of the seeds remains constant after drying, which is recorded as W 2 , thus calculate the seed water content (%) = (W 1 -W 2 ) / W 1 ×100%. Each treatment was repeated 5 times, and the average value was taken.

[0053] 2. Result analysis

[0054] Table 1 shows the water content of seeds

[0055] repeat

[0056] It can be seen from the table that the thousand-grain weight of garlic with ribbed leaves is 0.4924g, and the average water content is 8.328%.

Embodiment 3

[0057] Example 3 A method for extracting genomic DNA from leaves of wild relatives of garlic, slightly modified with reference to the method of Shi C, including the following steps:

[0058] (1) Use the dried leaves of the wild relative species of garlic (Allium prismatica) as materials, pick fresh leaves during the leaf-expansion stage, put them in a ziplock bag with silica gel, and dry them for 15-30 days.

[0059] (2) Weigh 2 g of completely dried leaves, grind them into powder quickly under liquid nitrogen conditions, and transfer them to a centrifuge tube.

[0060] (3) Add 65°C preheated SDS extraction buffer (same as in Example 1) to the tube, add 20 μL of precooled β-hydroxyethanol, and quickly vortex to mix.

[0061] (4) Place the centrifuge tube in a 65°C water bath and keep it warm for 60 minutes, during which time it is mixed slowly 8 times.

[0062] (5) Take out the centrifuge tube and cool it to room temperature, wipe off the water droplets on the outer wall of t...

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Abstract

The invention belongs to the field of biotechnology, and discloses a method for extracting DNA from Xinjiang garlic wild relative species. The method mainly comprises: collecting mature seeds in a primary environment, and naturally drying, wherein the same batch of the seeds achieving the safe water content are used for extracting DNA; placing the seeds in a mortar, rapidly grinding into powder, adding an extraction liquid, placing into a water bath pot, leaching, and carrying out centrifugation; taking the supernatant, adding chloroform-isoamyl alcohol, and carrying out centrifugation; takingthe supernatant, adding chloroform-isoamyl alcohol again, and carrying out centrifugation; taking the supernatant, adding pre-cooled isopropanol, precipitating, and carrying out centrifugation to obtain a precipitate; and discarding the supernatant, adding ethanol, rinsing the precipitate, removing the supernatant, carrying out natural air-drying, and adding double distilled water to completely dissolve the precipitate so as to obtain the DNA extraction raw liquid. According to the present invention, the accurate identification is achieved during the taking of the Xinjiang garlic wild relative species, the work is safe, and the efficiency is high; and the DNA extraction operation process has the low coat and is rapid, and the interference caused by sulfur-containing compounds and other components can be effectively reduced so as to obtain the high-quality seed genomic DNA.

Description

technical field [0001] The invention relates to the technical field of plant molecular biology research, in particular to a method for extracting high-quality seed genome DNA of wild relatives of garlic. Background technique [0002] Allium plants include garlic, leek, onion, onion and other cultivated species. Xinjiang is located in Central Asia. The Tianshan Mountains, Altai Mountains, Kunlun Mountains, Junggar Basin and Tarim Basin constitute the special topography and landform of this area, and the geographical environment is complex. Diverse, unique floristic composition, germplasm resources are different from Northeast China, North China, Southwest China and other provinces in Northwest China, and contain wild relatives such as garlic, leek, onion, onion, including wild onion, wild garlic and wild leek Among them, the wild garlic distributed in the wild has long been used by local herdsmen as vegetables, medicinal and ornamental plants. In China, it is only distributed...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 林辰壹马晓蓓叶远荣
Owner XINJIANG AGRI UNIV
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