Method for efficiently extracting solanaceae seed RNA (Ribonucleic Acid)

A technology of solanaceae and seeds, applied in the field of RNA extraction, can solve the problems of many operation steps, long extraction time, high cost, etc., and achieve the effect of high yield, strong practicability and reliability, and easy operation

Inactive Publication Date: 2013-05-01
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These methods are either expensive, or have

Method used

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  • Method for efficiently extracting solanaceae seed RNA (Ribonucleic Acid)
  • Method for efficiently extracting solanaceae seed RNA (Ribonucleic Acid)
  • Method for efficiently extracting solanaceae seed RNA (Ribonucleic Acid)

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Embodiment 1

[0021] Embodiment 1: the method for efficiently extracting tomato fresh seed RNA, concrete operations are as follows:

[0022] (1) Take fresh tomato seeds, dried tomato seeds, fresh capsicum seeds, and dried capsicum seeds, respectively. Soak these four materials in sodium hypochlorite with a mass percentage concentration of 10% for 15 minutes, and wash them with sterilized water for 3 times. , cut the seeds, discard the endosperm, take 18 halves (100mg) and grind them into powder with liquid nitrogen;

[0023] (2) Add 100ul sterilized extract solution (1% Saracosyl, 100mM Tris-HCl, 280mM NaCl, 10mM EDTA, 10ul / ml β-mercaptoethanol, pH 8.5) to the ground sample, then add 100ul phenol, fully Vortex, place in ice-water mixture for 10 min, centrifuge at 10,000 g for 5 min at 4°C;

[0024] (3) After centrifugation, draw the supernatant into a 1.5ml centrifuge tube, add 500ul chloroform / isoamyl alcohol mixture to the supernatant according to the ratio of 500ul chloroform / isoamyl al...

Embodiment 2

[0031] Embodiment 2: the method for efficiently extracting tomato fresh seed RNA, concrete operations are as follows:

[0032] (1) Take fresh tomato seed material, soak it in sodium hypochlorite with a mass percentage concentration of 15% for 10 minutes, wash it with sterilized water for 4 times, cut the seed open, discard the endosperm, add liquid nitrogen and grind it into a powder;

[0033] (2) Add 110ul sterilized extract solution (1% Saracosyl, 100mM Tris-HCl, 280mM NaCl, 10mM EDTA, 10ul / ml β-mercaptoethanol, pH 8.5) to 80mg of ground sample, then add 110ul phenol, Vortex thoroughly, place in ice-water mixture for 15 min, centrifuge at 10000g for 5 min at 4°C;

[0034] (3) After centrifugation, draw the supernatant into a 1.5ml centrifuge tube, and add chloroform / isoamyl alcohol to the supernatant according to the ratio of 600ul of chloroform / isoamyl alcohol mixture per ml of supernatant. solution (volume ratio 23:1), vortex fully, centrifuge at 10000g for 5min at 4°C, ...

Embodiment 3

[0036] Embodiment 3: the method for efficiently extracting capsicum dry seed RNA, concrete operations are as follows:

[0037] (1) Take the dried chili seed material, soak it in sodium hypochlorite with a mass percentage concentration of 12% for 12 minutes, wash it with sterilized water for 3 times, cut the seed, discard the endosperm, add liquid nitrogen and grind it into a powder;

[0038] (2) Add 120ul sterilized extract (1% Saracosyl, 100mM Tris-HCl, 280mM NaCl, 10mM EDTA, 10ul / ml β-mercaptoethanol, pH 8.5) to 90mg of ground sample, then add 120ul phenol, Vortex fully, place in ice-water mixture for 20 min, centrifuge at 10000g for 5 min at 4°C;

[0039] (3) After centrifugation, draw the supernatant into a 1.5ml centrifuge tube, add 550ul chloroform / isoamyl alcohol mixture to the supernatant according to the ratio of adding 550ul chloroform / isoamyl alcohol to the supernatant and mix solution (volume ratio 25:1), vortex fully, centrifuge at 10000g for 5min at 4°C, the so...

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Abstract

The invention discloses a method for efficiently extracting solanaceae seed RNA (Ribonucleic Acid). The method has the characteristics of low difficulty in operation steps of the RNA extraction and high efficiency, and can be used for extracting not only fresh seeds of solanaceae plants such as tomato but also dry seeds thereof; the problems of over-high protein residue and serious RNA degradation during extraction of the solanaceae seeds by the conventional method are solved; and the method can be directly used for extracting the RNA of the solanaceae seeds and has the advantages of easiness in mastering of the technology and high application performance.

Description

technical field [0001] The invention belongs to the field of molecular biology, in particular to a method for extracting RNA from fresh or dry seeds of Solanaceae plants. Background technique [0002] 70% of human food comes directly from plant seeds, which are also the main carrier of plant genetic resources. In the fields of seed formation, germination, and seedling establishment, molecular biology research based on RNA molecules, such as gene dynamic expression, cDNA library construction, gene cloning, and in vitro translation, requires sufficient and high-quality RNA. Rapid RNA extraction methods become a key technology. [0003] The types and quantities of RNA molecules have distinct time and space characteristics, and specific research materials must often be used in research. Polysaccharides, proteins, fats, and some secondary metabolites contained in plant materials can largely interfere with the extraction quality and recovery efficiency of RNA. The composition o...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 韩芹芹高凯宋玉竹张金阳
Owner KUNMING UNIV OF SCI & TECH
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