Simplified method for extracting RNA of polyphenol plants based on nano magnetic beads

A nano-magnetic bead and extraction method technology, applied in the biological field, can solve the problems of large differences in the quality of extracted samples, high requirements for the operating environment, and the physical and mental impact of operators, so as to eliminate discomfort and environmental pollution and improve test efficiency. , the effect of short operation time

Active Publication Date: 2017-12-29
HENAN UNIV OF ANIMAL HUSBANDRY & ECONOMY
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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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  • Simplified method for extracting RNA of polyphenol plants based on nano magnetic beads
  • Simplified method for extracting RNA of polyphenol plants based on nano magnetic beads
  • Simplified method for extracting RNA of polyphenol plants based on nano magnetic beads

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Embodiment Construction

[0038] The simplified extraction method of polyphenolic plant RNA based on nano magnetic beads of the present invention comprises the following steps:

[0039] (1) Sample preparation: Take 0.2 g of polyphenolic plant tissue material and put it into a pre-cooled mortar, add excess liquid nitrogen, and quickly grind it into a uniform powder. During the grinding process, ensure that the tissue material is always immersed in liquid nitrogen;

[0040] (2) Cell lysis: Before the liquid nitrogen is completely volatilized, transfer the material into a 1.5ml centrifuge tube, then add 300uL lysate and 20uL nano-magnetic beads in sequence, pipette and mix the mixed solution for 5 times, and then static at room temperature at 25°C. set for 10min;

[0041] (3) Washing: transfer the 1.5ml centrifuge tube to the magnetic separation rack, let it stand for 1 min, discard the supernatant with a pipette gun, and then remove the magnetic separation rack. Then add 600uL of washing solution, pipe...

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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...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 陈黄曌邓晓旭卜文静余海尤侯文静琚瑾李澎瀛张良玉王梦迪
Owner HENAN UNIV OF ANIMAL HUSBANDRY & ECONOMY
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