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Hermatypic coral total RNA extraction method

An extraction method, coral technology, applied in the direction of DNA preparation, recombinant DNA technology, etc., to achieve the effects of clearly visible bands, reducing loss, and reducing the loss of RNA amount in reef-building corals

Inactive Publication Date: 2016-03-23
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to solve the problem that there is no RNA extraction method suitable for reef-building corals in the prior art, and to provide a method for extracting the total RNA of reef-building corals; the RNA of reef-building corals extracted by this method has high purity and good quality. It greatly reduces the RNA loss and DNA pollution of reef-building corals, and can be applied to the requirements of molecular biology experiments such as subsequent RT-PCR

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  • Hermatypic coral total RNA extraction method
  • Hermatypic coral total RNA extraction method
  • Hermatypic coral total RNA extraction method

Examples

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Embodiment 1

[0025] In July 2011, I dived in the waters of Sanya Bay to collect samples of Pocillopora staghorns, clustered helmet-shaped corals and Porites orange. The seawater depth is 1-5m. The samples were stored with liquid nitrogen to ensure that the materials can be frozen at ultra-low temperature quickly. The above three reef-building coral species were identified by Huang Hui, a researcher at the South China Sea Institute of Oceanology, Chinese Academy of Sciences.

[0026] Use high temperature sterilization and 0.22μ microporous filter seawater to wash the Pocillum staghorn coral for 2-3 times to remove other attached organisms on the surface of the Pocillum staghorn coral; After the water on the surface of the coral was sucked dry, about 3g of staghorn cup coral tissue was weighed and ground into powder in a mortar filled with liquid nitrogen. Transfer the ground and crushed Pocillopora staghorn tissue powder to a clean EP tube, then add 1 mL of guanidine isothiocyanate lysate, ...

Embodiment 2

[0036] In July 2011, I dived in the waters of Daya Bay and Sanya Bay to collect samples of Frost Staghorn Coral and Porites Porites. The depth of the seawater was 1-5m. The samples were stored on dry ice during sampling. For long-term storage, they should be frozen in a -80°C ultra-low temperature refrigerator. The above two species of reef-building stony corals were identified by Huang Hui, a researcher at the South China Sea Institute of Oceanology, Chinese Academy of Sciences.

[0037] Rinse Frost Acropora repeatedly with high temperature sterilized seawater filtered through a 0.22μ microporous membrane to remove other attached organisms on the surface of Frost Staghorn Coral; pre-cool the mortar and dry the water on the surface of Frost Staghorn Coral Finally, weigh about 3g of staghorn coral tissue and grind it into powder in a mortar filled with liquid nitrogen. Transfer the ground and crushed Frost Acropora tissue powder to a clean EP tube, then add 1 mL of guanidine is...

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Abstract

The invention discloses a hermatypic coral total RNA extraction method. The extraction method comprises the following steps: washing the surface of a hermatypic coral sample, which is freshly collected and is preserved at an ultralow temperature, by sterile seawater to remove other organisms adhered on the surface of the sample, then grinding the sample in liquid nitrogen, cracking the sample by an isothiocyanic acid cracking liquid so as to release the free hermatypic coral RNA to the isothiocyanic acid cracking liquid, and then extracting the hermatypic coral total RNA; wherein the isothiocyanic acid cracking liquid contains the following components: 38 vol.% of acidic saturated phenol, 0.8M of guanidinium thiocyanate, 0.4M of ammonium thiocyanate, 0.1M of sodium acetate, 5 vol.% of glycerin, and the balance being RNase-free water. The provided method can well extract hermatypic coral total RNA, maximally reduces the degradation of hermatypic coral total RNA, and can be applied to various molecular biological researches such as hermatypic coral gene clone, differential display analysis, molecular hybridization, and the like.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a method for extracting total RNA of reef-building stony corals. Background technique [0002] Coral reefs are one of the ecosystems with the highest biodiversity on the earth. Coral reef ecosystems not only provide various biological resources for human production and life, but also have huge ecological functions and ecological values. Balance plays an important role. However, in recent years, the phenomenon of coral bleaching and death has become more and more frequent around the world, leading to the degradation of coral reefs in severe cases. Studying the mechanism of coral bleaching requires not only understanding the external physiological responses of corals, but also a deep understanding of the internal molecular mechanisms that cause these changes. There are many types of reef-building corals, with more than 70 genera. Obtaining high-quality total RNA ...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 胡思敏许昌有黄晖刘胜
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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