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Method for extracting RNA (Ribonucleic Acid) of eremophyte Reaumuria songarica Maxim

An extraction method, a technology of desert plants, applied in the direction of DNA preparation, recombinant DNA technology, etc., can solve the problems of many operation steps, long time-consuming, low yield, etc., and achieve the goal of simplifying the operation process, reducing unnecessary losses and high yield Effect

Inactive Publication Date: 2012-10-31
COLD & ARID REGIONS ENVIRONMENTAL & ENG RES INST CHINESE
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  • Application Information

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Problems solved by technology

This method can completely remove pollutants such as DNA and increase the yield of total RNA. However, this method has relatively many steps, takes too long, and the yield is relatively low.

Method used

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  • Method for extracting RNA (Ribonucleic Acid) of eremophyte Reaumuria songarica Maxim
  • Method for extracting RNA (Ribonucleic Acid) of eremophyte Reaumuria songarica Maxim
  • Method for extracting RNA (Ribonucleic Acid) of eremophyte Reaumuria songarica Maxim

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Embodiment Construction

[0027] The technical solution of the present invention will be further described below in conjunction with the accompanying drawings.

[0028] The specific steps of the test technology of desert plant red sand high-quality total RNA extraction are as follows:

[0029] 1. Test preparation stage:

[0030] 1. Plant material

[0031] The red sand with uniform size and vigorous growth in the desert of Hexi Corridor was selected, and the leaves were immediately frozen in liquid nitrogen and stored at -80°C for later use.

[0032] 2. Reagents

[0033] Sodium dodecyl sulfate (SDS), cetyltrimethylammonium bromide (CTAB), trishydroxymethylaminomethane (Tris), ethylenediamine tetraacetate (EDTA), diethyl pyrocarbonate (DEPC ), b-mercaptoethanol, sodium acetate, and lithium chloride were purchased from American Amresco Company, RNase-free RQ DNase enzyme and cross-linked polyvinylpyrrolidone (PVPP P 6755) were purchased from American Sigma Company, Taq enzyme (5 U / μL ), dNTP (2.5 mmol...

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Abstract

The invention relates to a method for extracting RNA (Ribonucleic Acid) of eremophyte Reaumuria songarica Maxim. The method comprises the following steps of: scattering PVPP (Crosslinked Polyvinyl Pyrrolidone) to leaves of Reaumuria songarica Maxim, grinding, centrifuging an obtained solution, taking supernate, adding chloroform and isoamylol which are equal in volume, and centrifuging; taking supernate again, adding phenol, chloroform and isoamylol which are equal in volume, and centrifuging; taking supernate, adding chloroform and isoamylol which are equal in volume, and centrifuging again; taking supernate, adding precooled absolute ethyl alcohol and LiC1, mixing uniformly, and performing ice bath; centrifuging again, pouring supernate, adding precooled absolute alcohol and sodium acetate, uniformly mixing, placing in a refrigerator, and centrifuging; and abandoning supernate, rinsing twice with 75% ethyl alcohol, centrifuging, collecting precipitates, airing to obtain a pure RNA sample, and dissolving the RNA sample in DEPC (Diethylpyrocarbonate) water. According to the invention, by means of effectively removing polyphenol, polysaccharide, protein, ion substances and other secondary metabolite, the pure RNA sample is obtained; and in addition, the yield is high and less time is consumed. Compared with the prior art, the method disclosed by the invention is simple to operate, and RNA is not degraded by RNase (Ribonuclease) and is good in integrity.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a method for extracting desert plant red sand RNA. Background technique [0002] The strong stress resistance of desert plants has always attracted the attention of scholars. This strong stress resistance is mainly due to the large amount of polysaccharides, polyphenols, tannic acid and other secondary metabolites in desert plants. The application of modern molecular biology techniques plays an important role in revealing the strong resistance mechanism of desert plants and the protection of genetic germplasm resources. Detecting changes in gene expression levels under different environmental stresses provides some necessary basic data for understanding plant stress resistance. The extraction of high-quality RNA is a necessary prerequisite for gene expression research in molecular biology. There are many methods for extracting RNA, including guanidine isothiocyanate ...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 王小华肖洪浪肖生春赵亮邹松兵
Owner COLD & ARID REGIONS ENVIRONMENTAL & ENG RES INST CHINESE
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