Universal type total RNA extraction kit and method

A general-purpose, kit-based technology used in the field of molecular biology to solve problems such as unsatisfactory results

Active Publication Date: 2017-10-20
北京爱普拜生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when we use these methods to extract RNA from plant fruits, we still cannot obtain satisfactory results

Method used

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  • Universal type total RNA extraction kit and method
  • Universal type total RNA extraction kit and method
  • Universal type total RNA extraction kit and method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] 1. Experimental method

[0085] Grind barley leaves, corn leaves, rice leaves (Nipponitril), sorghum leaves, soybean leaves, paulownia leaves, tomato leaves, dandelion leaves, and pothos leaves into powder in liquid nitrogen.

[0086] Take 0.05g of the powder and place it in a centrifuge tube with 1mL of RAP1 lysate, vortex and mix well, and place at room temperature for 10 minutes, during which time vortex 3-4 times.

[0087] Add 200 μL of chloroform, vortex to mix, and place at room temperature for 10 minutes. Vortex 3-4 times in between. Centrifuge at 13000 rpm at 4°C for 15 minutes.

[0088] Aspirate 500 μL of supernatant, add 500 μL of isopropanol, and mix gently.

[0089] Add the mixture (solution and possible flocculent precipitate) obtained in the previous step to an adsorption column RB (the RNA adsorption column is placed in the collection tube), centrifuge at 12,000rpm for 30 seconds, pour off the waste liquid, and put the adsorption column RB into the col...

Embodiment 2

[0100] 1. Experimental method

[0101] Take 0.05 g of rice leaves, rice roots, rice stems, dandelion flowers, and tomato fruit powders and place them in a centrifuge tube with 1 mL of RAP1 lysate, vortex and mix, and place at room temperature for 10 minutes, vortexing 3-4 times during the process.

[0102] Add 200 μL of chloroform, vortex to mix, and place at room temperature for 10 minutes. Vortex 3-4 times in between. Centrifuge at 13000 rpm at 4°C for 15 minutes.

[0103] Aspirate 500 μL of supernatant, add 500 μL of isopropanol, and mix gently.

[0104] Add the mixture (solution and possible flocculent precipitate) obtained in the previous step to an adsorption column RB (the RNA adsorption column is placed in the collection tube), centrifuge at 12,000rpm for 30 seconds, pour off the waste liquid, and put the adsorption column RB into the collection tube. tube;

[0105] Add 700 μL of RAW washing solution to the adsorption column RB, centrifuge at 12,000 rpm for 30 seco...

Embodiment 3

[0115] 1. Experimental method

[0116] Take yak lung, yak liver, mouse liver tissue, mouse lung tissue, rat pancreas tissue, rat parotid gland, pork, and mutton tissue and grind them into powder in liquid nitrogen. In a centrifuge tube, vortex to mix, and place at room temperature for 10 minutes, during which time vortex 34 times.

[0117] Add 200 μL of chloroform, vortex to mix, and place at room temperature for 10 minutes. Vortex 3-4 times in between. Centrifuge at 13000 rpm at 4°C for 15 minutes.

[0118] Aspirate 500 μL of supernatant, add 500 μL of isopropanol, and mix gently.

[0119] Add the mixture (solution and possible flocculent precipitate) obtained in the previous step to an adsorption column RB (the RNA adsorption column is placed in the collection tube), centrifuge at 12,000rpm for 30 seconds, pour off the waste liquid, and put the adsorption column RB into the collection tube. tube;

[0120] Add 700 μL of RAW washing solution to the adsorption column RB, c...

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Abstract

The invention belongs to the field of molecular biology and mainly relates to a simple, efficient and universal type total RNA extraction kit and method. The kit is applicable to extraction of high-purity, high-integrity and high-yield total RNA from a majority of biological samples such as plant samples, animal samples, microorganism samples, blood samples as well as different tissue parts including roots, stems, leaves, flowers and fruits of plants, has the advantages of rapidness, convenience, efficiency, universality and the like, and pigments, grease and other impurities in the RNA extraction process can be simply and conveniently removed to the maximum extent due to a thin adsorption column. The extracted total RNA can be directly used for inverse transcription and other downstream molecular biology experiment operations.

Description

technical field [0001] The invention patent belongs to the field of molecular biology. The invention relates to a simple and efficient universal total RNA extraction kit and method. This kit and method are suitable for extracting high-purity, high-integrity, and high-yield total RNA from most biological samples such as plants, animals, microorganisms, and blood, as well as plant roots, stems, leaves, flowers, and fruits. Total RNA can be directly used for reverse transcription and other downstream molecular biology experiments. Background technique [0002] With the continuous emergence of transcriptomics, proteomics, metabolomics and other omics, biological research has entered the post-genome era. As a first-developed technology, transcriptomics has been widely used in frontier biological research. Applications. Transcriptomics (transcriptomics) is a subject that studies gene transcription and transcriptional regulation in cells at the overall level. Simply put, transc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/101C12Q2523/308C12Q2523/113
Inventor 于祥春冯晓燕林挺王文利龚建
Owner 北京爱普拜生物技术有限公司
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