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Plant mRNA (messenger ribonucleic acid) extraction method

An extraction method and plant technology, applied in the field of molecular biology, acupuncture needles to extract mRNA from plant materials, can solve the problems of complicated extraction of mRNA, damage to plant materials, low efficiency, etc., and achieve strong practicability, wide application, Ease of operation

Pending 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] In order to solve the problems of complex operation, low efficiency, and damage to plant materials by traditional methods of extracting mRNA, a plant mRNA extraction method is provided.

Method used

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  • Plant mRNA (messenger ribonucleic acid) extraction method

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specific Embodiment approach 1

[0014] Specific embodiment one: the plant mRNA extraction method of the present embodiment carries out according to the following steps:

[0015] 1. Treatment of acupuncture needles: put the acupuncture needles into a brown bottle, and in an ultrasonic cleaner, use hexane, acetone and ethanol solutions to vibrate and clean the acupuncture needles successively; then put the brown bottle in an oven at 75°C to 82°C to dry ;

[0016] 2. Incubate the dried acupuncture needles in a mixed solution of trimethoxysilane, xylene and diisopropylethylamine, and incubate at 75°C to 82°C for 10h to 16h; use nitrogen to isolate oxygen during the operation , to prevent the oxidation of the medicine; the volume ratio of the diisopropylethylamine, xylene and trimethoxysilane is 1: (70-80): (20-30);

[0017] 3. Wash the cultured acupuncture needles with 48℃~52℃ ethyl acetate solution for 3 to 5 times, then put the acupuncture needles into the mixed solution of 0.1MKOH and 0.1-0.5μg / ml polythymid...

specific Embodiment approach 2

[0020] Embodiment 2: The difference between this embodiment and Embodiment 1 is that in Step 1, the time for vibrating and cleaning the acupuncture needles is 0.2 hours to 2 hours. Other steps and parameters are the same as those in the first embodiment.

specific Embodiment approach 3

[0021] Embodiment 3: The difference between this embodiment and Embodiment 1 is that the drying time of Step 1 is 0.2h-2h. Other steps and parameters are the same as those in the first embodiment.

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Abstract

The invention discloses a plant mRNA (messenger ribonucleic acid) extraction method, belongs to the technical field of molecular biology, and particularly relates to the technical field of extracting mRNA from plant materials through acupuncture needles.The plant mRNA extraction method is provided to solve the problems of operation complexity, low efficiency and damage to plant materials in conventional mRNA extraction methods.The plant mRNA extraction method includes 1, treating an acupuncture needle; 2, putting the dried acupuncture needle into a mixed solution of trimethoxysilane, xylene and diisopropylethylamine for incubation; 3, embedding the acupuncture needle to enable poly-thymine to be adsorbed to the surface of the acupuncture needle; 4, autoclaving the embedded acupuncture needle subjected to ultrapure water elution; 5, punching the naturally dried acupuncture needle into a test position of a target gene of a tender plant material, keeping the acupuncture needle at the test position for 1-3 minutes, pulling out the acupuncture needle, and putting the acupuncture needle into a PCR (polymerase chain reaction) tube of diethyl pyrocarbonate water to complete plant mRNA extraction.The plant mRNA extraction method has the advantages that total RNA extraction is not needed, the mRNA can be extracted from the plant materials in 1-3 minutes, and simplicity, convenience and rapidness of the operation process are achieved.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, in particular to the technical field of extracting mRNA from plant materials with acupuncture needles. 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] In order to solve the problems of complex operation, low efficiency and damage to plant materials by the traditional method for extracting...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12Q1/68
CPCC12N15/1003C12Q1/6806
Inventor 王玲卡尔·哈森斯坦
Owner NORTHEAST FORESTRY UNIVERSITY
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