Method for extracting DNA (Deoxyribonucleic Acid) of total genome from zooplankter and intestinal inclusions thereof

A zooplankton and extraction method technology, applied in the direction of recombinant DNA technology, DNA preparation, etc., can solve the problem that there is no patent report on the total genome DNA extraction method of zooplankton and intestinal contents, and achieve the effect of reducing loss

Inactive Publication Date: 2011-11-23
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

[0004] At present, there is no patent report on the extraction method of total genomic DNA from zooplankton and intestinal contents

Method used

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  • Method for extracting DNA (Deoxyribonucleic Acid) of total genome from zooplankter and intestinal inclusions thereof
  • Method for extracting DNA (Deoxyribonucleic Acid) of total genome from zooplankter and intestinal inclusions thereof
  • Method for extracting DNA (Deoxyribonucleic Acid) of total genome from zooplankter and intestinal inclusions thereof

Examples

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Embodiment 1

[0027]In March 2011, the zooplankton was dragged horizontally in the Sanya coral reef sea area with a zooplankton I net, and after the zooplankton was fixed with neutral Luger's solution, the zooplankton was analyzed under a stereomicroscope with a disposable plastic straw. For selection, put the same species into 1.5ml sterilized centrifuge tubes, fix with neutral Luger's solution, and store at 4°C for later use. Take 20 fixed Eucalanus subcrassus and place them in a petri dish with a diameter of 3.5cm, wash the surface of zooplankton gently with sterilized seawater filtered through a 0.2μm Millipore membrane, and check with a microscope to ensure that the surface of the zooplankton is clean. No other organisms attached; take out the zooplankton, and then wash with sterilized ultrapure water to reduce the impact of salt ions in sterile filtered seawater on DNA extraction, transfer the zooplankton to a 1.5ml centrifuge tube, and blot the water dry. Freeze the centrifuge tube i...

Embodiment 2

[0033] In May 2011, the zooplankton was dragged horizontally in the Daya Bay sea area with a zooplankton type I net, and after the zooplankton was fixed with neutral Luger's solution, the zooplankton was sorted under a stereomicroscope with a disposable plastic straw , put the same species into 1.5ml sterilized centrifuge tubes, fix with neutral Luger's solution, and store at 4°C for later use. Take 20 fixed fat soft arrow worms (Flaccisagitta enflata) and place them in a petri dish with a diameter of 3.5 cm. Repeatedly and gently wash the zooplankton body surface with sterilized seawater filtered through a 0.2 μm Millipore membrane, and examine under a microscope to ensure that the body surface is No other organisms attached; take out the zooplankton, and then wash with sterilized ultrapure water to reduce the impact of salt ions in sterile filtered seawater on DNA extraction, transfer the zooplankton to a 1.5ml centrifuge tube, and blot the water dry. Freeze the centrifuge t...

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Abstract

The invention discloses a method for extracting DNA (Deoxyribonucleic Acid) of a total genome from zooplankter and intestinal inclusions thereof. The method comprises the following steps of: immobilizing the zooplankter by utilizing a neutral Ruge's solution so that the intestinal inclusions are not lost and then to washing the body surface by utilizing sterile seawater filtered by a filtering membrane of 0.2 micron to remove other living beings attached on the surface so that polluting the zooplankter and the DNA of the intestinal inclusions is avoided; then washing by utilizing sterile super-pure water to reduce influence on DNA extraction caused by the salt ions in seawater; crushing and then cracking by utilizing a DNA cracking solution to release and dissociate the DNA of the zooplankter and intestinal inclusions to the cracking solution; then extracting the DNA of the total genome from the zooplankter and the intestinal inclusions thereof through utilizing a regular method. By using the method provided by the invention, the extraction of the DNA of the total genome from the zooplankter and the intestinal inclusions thereof is realized, and the loss of the DNA amount of the intestinal inclusions of the zooplankter is reduced to the maximum extent; and high-quality DNA samples for accurately constructing a qualitative structure of a spot plankton food web are provided, and the function and the efficiency of a regionally ecological system are further analyzed.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for extracting total genome DNA of zooplankton and intestinal contents thereof. Background technique: [0002] Zooplankton is the secondary producer of the food chain, and it is in the pivotal position in the marine food web. It is the main driver of the marine biological pump and has a significant effect on the biogeochemical cycle of biogenic elements. Therefore, carrying out research on its feeding ecology is of great significance for understanding the structure and function of marine ecosystems, quantifying the cascading effects of matter and energy along the food chain, and exploring the level of biological productivity in sea areas and its changing laws. Using molecular biology techniques can break through the "cultivation" link, directly study the intestinal food composition and feeding efficiency of zooplankton in the natural environment, clarify its cor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 郭志灵刘胜林森杰李涛胡思敏
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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