Method for extracting hybrid total RNA for gene chips from animal dispose tissues

Abstract

The invention relates to a method for extracting hybrid total RNA for gene chips from animal dispose tissues, which comprises the following steps: 1) tissue grinding and Trizol lysis; 2) chloroform extraction: adding chloroform to lytic tissues to extract RNA; 3) secondary chloroform extraction: getting supernate obtained in step 2), and adding chloroform to the supernate for extraction; 4) isopropanol precipitation: taking the aqueous phase obtained in step 3), adding isopropanol with the same volume as that of the aqueous phase, and mixing the aqueous phase and the isopropanol uniformly; 5) total RNA washing: adding alcohol with the mass concentration of 75% to precipitates obtained in step 4), and blowing the precipitates to enable the precipitates to suspend; and 6) total RNA dissolving: drying the precipitates obtained in step 5) in a clean bench at room temperature to obtain total RNA precipitates, and dissolving the total RNA precipitates with RNase-free liquid. The method is convenient and rapid.

Description

technical field [0001] The invention belongs to a method for extracting total RNA from animal adipose tissue, in particular to a method for extracting total RNA samples in animal adipose tissue for gene chip research. Background technique [0002] Adipose tissue is not only the body's energy storage, but also the largest endocrine organ in the body. Animal adipose tissue has a proliferation of cells in the early postnatal period, but it is still dominated by the increase of cell volume, especially in the later period or adulthood, it is easy to accumulate in the body. Fat metabolism includes fat mobilization (lipolysis), body movement and fat storage (lipogenesis). The deposition of animal body fat is a state of balance between fat synthesis and catabolism. When the anabolism is enhanced or the catabolism is decreased, the original balance will be broken and the fat deposition will increase. When more fat is broken down than synthesized, body fat decreases. An in-depth u...

AI Technical Summary

Problems solved by technology

[0003] At present, the RNA extraction technology is very mature, and there are also a large number of literature reports on the successful extraction of high-quality RNA using various methods and commercial reagents. Although these methods and reagents have many advantages, such as simple methods and high-quality extracted RNA, when encountering When the abundance of tissue RNA is extremely low, and RNA products are often accompanied by genomic DNA contamination and other problems, it is often impossible to obtain ideal results according to conventional procedures; due to the specificity of tissues, adipose tissues are rich in lipids, Fewer cells, mature adipocytes contain various organelles, nuclei, and a large central lipid droplet unlike other cell types

And the RNA content per unit cell is relatively small, the amount of total RNA contained in each mg tissue sample is less than 0.05 μg, so the yield of the extracted RNA is low, which increases the difficulty of extracting RNA from adipose tissue

Moreover, the RNA in adipose tissue used for gene chips is generally extracted with kits, which are relatively expensive. The kits are purchased from abroad, and it is very inconvenient to purchase.

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