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Method for identifying marine nematodes by denatured gradient gel electrophoresis

A technique of denaturing gradient gel and marine nematodes, which is applied in the field of identification of marine nematodes by DGGE technology, and achieves the effect of good distinction and identification

Inactive Publication Date: 2016-10-26
LIAOCHENG UNIV
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Problems solved by technology

[0008] The technical problem to be solved by the present invention is to apply deformation gradient gel electrophoresis (DGGE) to the identification of nematodes, and use this technology to study the composition of the community diversity of nematodes. In order to achieve the above objectives, the present invention provides a Application of DGGE technology in the identification and diversity research of marine nematodes

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  • Method for identifying marine nematodes by denatured gradient gel electrophoresis

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Embodiment Construction

[0034] The present invention will be further explained below in conjunction with embodiment.

[0035] References: Diez B., Pedros-Alio C., Marsh T. L., et a1. Application of denaturing gradient gel electrophoresis (DGGE) to study the diversity of marine picoeukaryotic assemblages and comparison of DGGE with other molecular techniques. Applied and Environmental Microbiology, 670 (7): 2942-2951.

[0036] EXAMPLES The method for identifying marine nematodes using deformable gradient gel electrophoresis comprises the following steps:

[0037] (9) Sampling and indoor sorting

[0038] (10) Morphological identification

[0039] (11) Extraction of total DNA

[0040] (12) Design and optimization of PCR primers

[0041] (13) Agarose electrophoresis detection

[0042] (14) Preparation of denaturing gradient gel

[0043] (15) Optimization of denaturing gradient gel electrophoresis conditions

[0044] (16) Using denaturing gradient gel electrophoresis to identify and analyze marine ...

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Abstract

The invention discloses a method for identifying marine nematodes by denatured gradient gel electrophoresis. The method is characterized by comprising the following steps: (1) sampling and indoor sorting, (2) morphological identification, (3) extraction of total DNA, (4) designing and optimizing of PCR primers, (5) agarose electrophoresis detection, (6) preparation of denatured gradient gel, (7) optimization of denatured gradient gel electrophoresis conditions, and (8) identification and analysis to the marine nematodes by denatured gradient gel electrophoresis. The method has the advantages that a DGGE technology is used for identifying the marine nematodes, and is used for analyzing community composition of the marine nematodes; according to a characteristic map, the migration rate of related nematodes on the denatured gradient gel can be obtained, differentiation and identification can be carried out well, and for species which are difficult to identify, more than two pairs of primers are required to be used in a combined manner at times.

Description

technical field [0001] The invention relates to the technical field of species identification and community composition detection of marine nematodes in marine sediments, in particular to a method for using DGGE technology in identifying marine nematodes. Background technique [0002] The meiobenthos are a class of animals that can pass through the 0.5mm mesh screen, but are trapped by the 0.042mm mesh screen. Free-living nematodes are the most dominant group of meiobenthos. In most habitats, the number of marine nematodes can account for 70%-90% of the total number of meiofauna, and in some habitats it can reach more than 90%. The high species diversity and abundance of marine nematodes suggest that they are one of the most evolved groups in the ocean. The role of this species in marine sediments lies not only in its numerical and biomass dominance, but also in its relationship to other species in the food web. Furthermore, marine nematodes, with their characteristic repr...

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2565/125
Inventor 孙静
Owner LIAOCHENG UNIV
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