A Simplified Extraction Method of Polyphenolic Plant RNA Based on Nanomagnetic Beads

Abstract

A simplified method for extracting RNA of polyphenol plants based on nano magnetic beads comprises the steps of (1), preparing samples; (2) allowing cell lysis; (3) washing, and repeating washing; (4) dissociating. Total RNA in polyphenol plants is extracted by the method of the invention under high rate, and the total RNA has high purity and complete fragments and is directly applicable to subsequent detection. Nucleic acid extraction time is shorter than that of the use of common nano magnetic beads, the efficiency is higher, the operation is simple, automation is easier to implement, the reagent environment is friendly, and the safety is better.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a simplified extraction method of polyphenolic plant RNA based on nano magnetic beads. Background technique [0002] RNA, Ribonucleic Acid, can transmit genetic information and is an important medium for regulating protein synthesis. The effective function of any gene depends on the expression and regulation of RNA, so RNA plays an important role in the field of molecular biology research. Extracting RNA with high purity and good integrity from plant tissues is the key to exploring the mechanism of RNA regulation. High-quality RNA extraction is an important prerequisite for molecular biology experiments such as cDNA (complementary deoxyribonucleic acid) library construction, fluorescent quantitative PCR detection, Northern hybridization, etc. [0003] At present, RNA extraction mainly adopts the guanidine isothiocyanate-phenol-chloroform one-step method proposed by Chom...

AI Technical Summary

Problems solved by technology

Phenolic substances are easily oxidized to quinones, which are irreversibly combined with RNA during the extraction process, thus affecting the purification of RNA

Moreover, phenolic substances remaining in the sample are prone to oxidation and produce dark brown substances, so the RNA extracted by traditional methods often has a deep rust color, which affects subsequent molecular experiments.

[0004] Commonly used extraction methods have the following disadvantages: 1. Conventional extraction reagents contain more toxic organic solvents, which pollute the environment and have a certain impact on the operator's body and mind; 2. Conventional extraction methods (non-nano magnetic bead method ) The operation takes a long time, and the operating environment is very demanding. If you don’t pay attention, it will cause the degradation of RNA, and the result will fall short; 3. The quality difference between the samples extracted by the conventional extraction method (non-nano magnetic bead method) is relatively large, the process is complicated, and the degree of automation is low. 4. The current conventional nano-magnetic bead method has low extraction efficiency to plant tissues rich in phenols, and usually uses two kinds of lotions to wash; 5. The current conventional nano-magnetic bead method must remove the total After the RNA is separated from the magnetic beads, the subsequent fluorescent quantitative PCR reaction can be carried out

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