Method for extracting total RNA from millet seeds

A seed and millet technology, applied in the field of biochemistry, can solve the problems of poor RNA extraction quality of millet

Inactive Publication Date: 2019-05-17
SHANXI AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] For above-mentioned situation, the purpose of the present invention is to provide a kind of method for extracting the total RNA of millet se

Method used

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  • Method for extracting total RNA from millet seeds
  • Method for extracting total RNA from millet seeds

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Embodiment 1

[0019] Adopt the method of the present invention to extract RNA, concrete operation is as follows:

[0020] (1) Preparation of RNA extraction buffer components: the extraction buffer is prepared by adding Tris-HCl, PVP, and β-mercaptoethanol to water in sequence. The volume ratio of the above components is: 1M Tris-HCl: 10% PVP: β-mercaptoethanol: water = 5:5:1:39, the Tris-HCl is Tris-HCl with a pH of 9.0;

[0021] The preparation method takes the preparation of 100mL extraction buffer as an example: take 10ml of Tris-HCl with pH 9.0, add 10ml of 10% w / v PVP, 2 mL of β-mercaptoethanol, and then add 78ml of water;

[0022] (2) Grind mature and plump millet seeds in liquid nitrogen to form a fine powder, add 0.7 mL of the above RNA extraction buffer for every 0.1 g of millet seed fine powder, mix well and let stand for 15 min, then centrifuge at 12 000 g at 4 °C 10 minutes;

[0023] (3) Add 1 mL of RNAiso Plus to the supernatant, mix well and let stand at room temperature for...

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Abstract

The invention relates to a method for extracting total RNA from millet seeds. The method comprises the following steps of (1) adding 0.1 g of liquid nitrogen-milled millet seed fine powder to 0.7 mL of a RNA extraction buffer, wherein the extraction buffer is prepared from Tris-HCl, PVP and beta-mercaptoethanol added with water in turn; (2) taking a supernatant and adding 1 mL of RNAiso Plus; (3)taking the supernatant, adding chloroform and mixing evenly; (4) taking the supernatant, transferring to another novel centrifuge tube, and adding an equal volume of isopropanol; (5) discarding the supernatant, washing a precipitate with ethanol, drying at the room temperature, and dissolving in RNase-free water to obtain RNA extract liquid. The method improves the quality and concentration of theRNA, reduces protein and salt ion pollution, and solves the problem of poor quality of millet RNA extraction.

Description

technical field [0001] The invention relates to the technical field of biochemistry, in particular to a method for extracting total RNA from millet seeds. Background technique [0002] Millet is a small miscellaneous grain in northern China. The nutritional value of the grain is very high, rich in protein, fat and vitamins. With the change of people's lifestyle, the demand for millet in the market is increasing, and the maintenance and germination of seed vitality The emergence of seedlings determines the reproduction and viability of plant species and the establishment of ecosystems, and directly affects the yield of crops. Since seed germination, especially the germination of small seeds such as millet, is more susceptible to environmental stress, mechanical injury, and pests and diseases, seed germination is considered to be the most important and vulnerable stage in the plant life cycle, so the study of molecular components of seed germination The mechanism helps to inc...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 温银元王金荣许冰霞尹美强裴帅帅柯贞进赵娟王计平冯文新王玉国
Owner SHANXI AGRI UNIV
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