Method for quickly extracting RNA through animal and plant tissue ablation apparatus based on ultrasonic principle

Abstract

The invention discloses a method for quickly extracting RNA through an animal and plant tissue ablation apparatus based on ultrasonic principle. The method comprises the following steps of (1) preserving tissues or cells with an extracting solution A, then adding an extracting solution B to the tissues or cells preserved in the extracting solution A, and performing uniform mixing so as to obtain amixture; (2) when RNA is extracted from the tissues, performing ultrasonic breaking on the mixture; (3) when RNA is extracted from the cells, directly performing operations in the step (5); (4) reversing the mixture after ultrasonic breaking to achieve a uniform mixing state; (5) adding chloroform to the mixture obtained through extracting the RNA from the cells in the step (3) or the mixture obtained in the step (4), performing centrifugation, and taking upper-layer liquid; (6) adding isopropyl alcohol to the upper-layer liquid, and then performing centrifugation so as to obtain precipitate;(7) adding an ethanol solution to the precipitate, performing uniform mixing, performing centrifugation, and discarding liquid; and (8) finally adding water for dissolving. The tissue ablation apparatus is used for ablating the tissues, the rate is high, incomplete tissue breaking and interference of endogenous RNA enzymes are avoided, the extraction time is shortened, and the extraction efficiency of the RNA is improved.

Description

technical field [0001] The invention belongs to the technical field of biological preparations, and in particular relates to a method for rapidly extracting RNA by using an animal and plant tissue ablation instrument based on the principle of ultrasound. Background technique [0002] In modern molecular biology experiments and clinical molecular diagnosis, it is often necessary to isolate and purify RNA from tissues and cells, and the quality of RNA often affects the success or failure of molecular biology experiments such as cDNA library, RT-PCR and Northern Blot. [0003] The commonly used method is to first protect the tissue with a tissue protection solution, and then use the extraction solution to extract the RNA in the tissue or cells. Commonly used protection methods include liquid nitrogen protection and protection solution protection, and the commonly used protection solutions include RNA later reagents and RNA safety reagents. [0004] Among them, the liquid nitro...

AI Technical Summary

Problems solved by technology

The liquid nitrogen grinding method requires a large tissue block, and it is difficult to completely break up the tissue block; the homogenization method is difficult to completely inhibit the activity of endogenous RNase, and it is easy to cause endogenous degradation; the disadvantage of the repeated freeze-thaw method is that it is difficult to Thoroughly lyses large tissues, and endogenous RNase activity is difficult to inhibit

Therefore, the RNA extraction of animal-derived tissue blocks has difficulties such as complicated operation, incomplete fragmentation, and difficulty in inhibiting the activity of endogenous RNA, which makes it difficult to guarantee the quality and yield of RNA extraction.

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