Method for extracting total RNA from tamarix chinensis tissues

A technology of tissue and tamarisk, applied in the field of RNA extraction of desert plant tamarisk, can solve the problems of difficult to find, RNA degradation, cumbersome and other problems

Inactive Publication Date: 2016-08-03
COLD & ARID REGIONS ENVIRONMENTAL & ENG RES INST CHINESE
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] According to reports, the improved CTAB method can extract total RNA in Tamarix tissue, but it is cumbersome, takes a long time, and the yield is low; even due to repeated operations, it is easy to cause RNA degradation and pollution; the current RNA extraction kit There are many types, but it is difficult to find a commercial RNA extraction kit suitable for T

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  • Method for extracting total RNA from tamarix chinensis tissues
  • Method for extracting total RNA from tamarix chinensis tissues
  • Method for extracting total RNA from tamarix chinensis tissues

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Embodiment

[0046] Summary of methods for extracting intraspecific total RNA from roots, stems, leaves and flower tissues of Tamarix:

[0047] The content of the invention includes four parts: the required kit equipment, the exclusive processing process before extraction, the extraction operation steps and the quality detection of the extracted RNA. The specific implementation plan is as follows:

[0048] 1. Required reagents and equipment

[0049] 0.1% DEPC water, absolute ethanol, liquid nitrogen, β-mercaptoethanol, E.Z.N.A? PlantRNAKit kit;

[0050] Centrifuge tube, test tube rack, medicine spoon, mortar, sterilizer, aluminum parchment paper, sterile paper towel, 1000ul pipette, 200ul pipette, 100ul pipette, 50ul pipette, 1000ul gun with filter Tip, 200ul tip with filter, 100ul tip with filter, 50ul tip with filter;

[0051] 2. Proprietary processing before extraction

[0052] Before extraction, soak the centrifuge tubes, medicine spoons and mortars with 0.1% DEPC aqueous solution ...

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Abstract

The invention discloses a method for extracting total RNA from tamarix chinensis tissues. According to the method, the traditional RNA extraction method is improved through using a kit in combination with the addition of PVPP powder, so that the RNA extraction effect and quality of plant materials which are not easy to extract are improved. The main technical process of the method comprises the following steps: treatment before kit equipment extraction, detailed RNA extraction, and later RNA concentration and quality control and detection. By using the method, the total RNA extracted from the tamarix chinensis tissues is high in concentration, good in quality and free of protein and pollution from other impurities. According to the method, a reference is provided for the RNA extraction of plants rich in salts and polyphenols in deserts.

Description

technical field [0001] The invention relates to a method for extracting desert plant tamarisk RNA. Background technique [0002] Extracting high-quality RNA is a necessary prerequisite for molecular biology research such as Northern analysis, RT-PCR, cDNA library construction, DD-PCR analysis, and in vitro translation, and is an important link in the study of gene expression; desert plants have strong stress resistance The main reason is that it contains a large number of secondary metabolites, such as polysaccharides, polyphenols, tannins and other substances. After the cells are broken, these substances will interfere with the extraction of RNA in different ways, seriously affecting the quality and yield of the extracted RNA. ; Therefore, finding out the interference that can inhibit or reduce secondary metabolites is a major problem in realizing a simple and feasible RNA extraction method. [0003] Tamarix ramosissima is a shrub or small tree, 1-5m high; the branches are...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1003C12Q1/6806
Inventor 燕霞马小飞钱朝菊尹成亮殷恒霞石勇
Owner COLD & ARID REGIONS ENVIRONMENTAL & ENG RES INST CHINESE
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