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Direct plant mRNA (messenger ribonucleic acid) extraction method

An extraction method and plant technology are applied in the field of directly extracting plant mRNA, which can solve the problems of low efficiency and cumbersome process of extracting plant mRNA, and achieve the effects of wide applicability, strong practicability, and precise positioning and extraction.

Inactive Publication Date: 2016-07-27
NORTHEAST FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The purpose of the present invention is to solve the traditional method to extract plant mRNA process is loaded down with trivial details, the problem of low efficiency, provides the direct extraction method of plant mRNA

Method used

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  • Direct plant mRNA (messenger ribonucleic acid) extraction method

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specific Embodiment approach 1

[0012] Specific embodiment one: the plant mRNA direct extraction method of the present embodiment is carried out according to the following steps:

[0013] 1. Preparation before extraction: wash the intradermal needle with hexane, acetone and ethanol solution in sequence, and dry it in an oven at 75°C to 82°C; then stack 2 to 4 slides and put them on the glass In the petri dish, hang the needle tip of the dried intradermal needle outward on the side of the glass slide, cover the lid of the petri dish, put it in an oven at 75°C to 82°C, remove the lid of the petri dish, dry it, and take it out Cover the petri dish and cool naturally; take 0.1mL-0.3mL trimethoxysilane, 0.7mL-0.9mL xylene and 5μL-10μL diisopropylethylamine in a centrifuge tube, and then place them in a mixer Mix for 5s-10s to get the mixed solution; then take 8uL-12uL mixed solution into the PCR tube, immerse the head of the intradermal needle in the mixed solution, and cover the PCR tube; incubate the intraderma...

specific Embodiment approach 2

[0016] Specific embodiment 2: The difference between this embodiment and specific embodiment 1 is that in step 1, 0.2 mL of trimethoxysilane, 0.8 mL of xylene and 8 μL of diisopropylethylamine are placed in a centrifuge tube, and then mixed Mix on the mixer for 5 s to obtain a mixed solution. Other steps and parameters are the same as those in the first embodiment.

specific Embodiment approach 3

[0017] Embodiment 3: This embodiment is different from Embodiment 1 in that the volume ratio of the KOH solution to polythymidine is 99:1. Other steps and parameters are the same as those in the first embodiment.

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Abstract

The invention provides a direct plant mRNA (messenger ribonucleic acid) extraction method, belongs to the biomolecular level technology field, and particularly relates to a method for extracting plant mRNA directly.The direct plant mRNA extraction method is provided to solve the problems of process complexity and low efficiency of conventional plant mRNA extraction methods.A new technique of directly extracting solid-phase mRNA genes is provided, Oligo dT(15-25) is adsorbed on the head surface of an intradermal needle after a series of treatments including washing, incubating, culturing, embedding and drying of the intradermal needle, the treated intradermal needle is punched at a set position of a plant material to extract mRNA of the plant material.By the direct plant mRNA extraction method, plant mRNA extraction is realized, the mRNA can be extracted from the plant material in 2 minutes, and simplicity, convenience and rapidness of the operation process are achieved.

Description

technical field [0001] The invention belongs to the technical field of biomolecules, and in particular relates to a method for directly extracting plant mRNA. Background technique [0002] High-quality RNA is an important prerequisite for plant molecular biology research. Due to the complex and diverse components of plant tissue cells, the extraction methods of plant tissue RNA are diverse. Traditional methods are commonly used in current research, including guanidine isothiocyanate method, phenol-SDS method, CTAB method, Trizol method, etc., all of which extract plant total RNA under the premise of cell rupture, and the experimental needs The mRNA only accounts for 1% to 4% of the total RNA, and the extraction process is cumbersome, with high efficiency and low cost. Contents of the invention [0003] The object of the present invention is to provide a method for directly extracting plant mRNA in order to solve the problems of cumbersome and low efficiency in the process...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1006C12Q2525/173C12Q2523/308
Inventor 卡尔·哈森斯坦王玲
Owner NORTHEAST FORESTRY UNIVERSITY
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