Extraction method for total RNA of feed type tetraploid robinia pseudoacacia leaves

A tetraploid Robinia pseudoacacia extraction method technology, applied in the field of plant molecular biology, can solve the problems of inability to extract high-quality RNA, difficult to effectively remove or inhibit, and unstable products, so as to reduce operating time and material consumption, Save operating time and cost, and easy to operate

Inactive Publication Date: 2012-04-25
NORTHEAST FORESTRY UNIVERSITY
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Problems solved by technology

Experiments have shown that using conventional equipment and common chemical reagents, it is difficult to effectively remove or inhibit the interference of substances such as proteins in the broken cells of Robinia pseudoacacia leav

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  • Extraction method for total RNA of feed type tetraploid robinia pseudoacacia leaves
  • Extraction method for total RNA of feed type tetraploid robinia pseudoacacia leaves
  • Extraction method for total RNA of feed type tetraploid robinia pseudoacacia leaves

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Embodiment Construction

[0026] The general idea of ​​the present invention is: based on common equipment and domestic chemical reagents, with the goal of eliminating the interference of protein and polysaccharide, it adopts the method of adding bentonite in the extract, adding a large amount of potassium acetate before precipitation, etc. to remove excessive protein and polysaccharide The key technology to obtain complete and stable RNA products. The specific method is: directly grind the leaves of Robinia pseudoacacia into powder in a centrifuge tube, add ice-bath buffer and β-mercaptoethanol, and centrifuge; discard the supernatant, add extraction buffer and β-mercaptoethanol, oscillate and mix, heat Water bath treatment; add water-saturated phenol and chloroform, oscillate to mix, and centrifuge; absorb the supernatant, add anhydrous ethanol and potassium acetate solution to mix, put in an ice bath and then centrifuge, discard the supernatant, and wash with 70% ethanol After centrifugation, the su...

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Abstract

The present invention relates to the field of plant molecular biology, specifically to an extraction method for total RNA of feed type tetraploid robinia pseudoacacia leaves. The extraction method is characterized by: filling tetraploid robinia pseudoacacia leaves in a centrifuge tube to carry out grinding; adding an ice bath buffer and beta-mercaptoethanol; centrifugating and removing supernatant; adding a CTAB extracting solution and beta-mercaptoethanol; centrifugating and taking supernatant; adding phenol and chloroform; uniformly mixing, centrifugating and taking supernatant; adding ethanol and a potassium acetate solution, uniformly mixing; carrying out ice bath, standing, and removing supernatant; cleaning by ethanol, and removing supernatant; carrying out air dry; adopting ddH2O to dissolve the RNA precipitation. The extraction method of the present invention has the following advantages that: the leaves are directly grinded in the centrifuge tube and are only subjected to extraction one time by the chloroform so as to reduce the operation time and the use amount of the material; bentonite is added to the extracting solution so as to well remove the protein interference; large amount of the potassium acetate is added to remove the polysaccharide so as to obtain the complete and stable RNA with high purity and high yield; the method has characteristics of simple operation, low cost, only requirements of ordinary equipment and domestic chemical reagents, and the like; the condition is provided for further molecular biology researches of the feed type robinia pseudoacacia and similar plants.

Description

technical field [0001] The invention relates to the technical field of plant molecular biology, in particular to a method for extracting total RNA from leaves of forage-type tetraploid Robinia pseudoacacia. Background technique [0002] Robinia genus (Leguminosae) Papilionaceae (Papilionaceae) plant, its leaves are rich in crude protein, sugar, minerals and vitamins, is a new type of feed with promising development prospects, especially the leaves of tetraploid Robinia. The content of protein and sugar is more abundant. In-depth study of the biological characteristics of feed-type tetraploid Robinia pseudoacacia is of great significance for the development of high-quality feed resources and the improvement of the breeding industry. However, because the forage-type tetraploid Robinia pseudoacacia leaves are rich in protein and polysaccharides, the extraction of total RNA is very difficult, which limits the research on the molecular biology of the forage-type tetraploid Robini...

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Application Information

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IPC IPC(8): C12N15/10
Inventor 刘磊孟凡娟黄凤兰
Owner NORTHEAST FORESTRY UNIVERSITY
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