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Method for sequencing partial DNA of mitochondrial Cytb gene of brachionus rotifer and a method for identifying brachionus rotifer

A DNA sequencing and DNA sequence technology, applied in the biological field, can solve the problems of inconclusive evolutionary path, insufficient morphological data to satisfy rotifer species identification and phylogenetic research, lack of in-depth understanding of ultrastructural and biochemical properties, etc. To achieve the effect of reducing species competition

Inactive Publication Date: 2019-02-19
ANHUI NORMAL UNIV
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Problems solved by technology

Early studies on rotifers focused on morphology and ultrastructure. Due to the variety of rotifer shapes and ecological habits, and the lack of in-depth understanding of its ultrastructure and biochemical properties, its evolutionary pathway has been difficult to determine. With the in-depth research on rotifers, the morphological data can no longer satisfy the identification and phylogenetic research of some species of rotifers.

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Embodiment Construction

[0013] The biological materials used in this method for identifying the genus Brachionus are Brachionus red, Brachiona square, Brachiona falciparum, Brachiona rectangle, Brachiona scissors, Brachiona split Rotifers, single individuals of each type of rotifers were picked and cultured as monoclonals. Among them, the formula of Gilbert (1963) was used for the red arm rotifers and the square arm rotifers. The ambient temperature is basically the same as that at the time of collection. The food used is Scenedesmus obliquus in the exponential growth phase cultured in HB-4 culture medium. The rotifers were filtered out, washed with sterile distilled water, collected in Eppendof tubes, and stored in a -20°C refrigerator.

[0014] Total DNA extraction:

[0015] Rotifer DNA was extracted using WizardTM Genomic DNA Purification Kit (pormega, USA). Take the frozen rotifers out of the -20°C refrigerator, centrifuge for a few seconds, and absorb excess water. Add 60 μl of nuclear lysis ...

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Abstract

The invention discloses a method for sequencing partial DNA of a mitochondrial Cytb gene of a brachionus rotifer and a method for identifying the brachionus rotifer, which comprises the following steps: extraction of DNA from the brachionus rotifer; PCR primer amplification and sequencing, wherein the PCR primer is Cytb1S: 5'-TWGATYTWCCTTCTCCHGC-3'; Cytb1R:5'-GGWGAYGCWGGACAD CTHCC-3; system analysis. The PCR primers can amplify part of the DNA of the mitochondrial Cytb gene, and the results can be used for precise identification of the rotifer, the identification problem caused by using morphology and ultrastructure is solved. In the process of large-scale culture of the rotifers, the competition of rotifers caused by mixed populations, which causes output reduction and large-scale death of the rotifers, can be reduced.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a partial DNA sequencing method of the mitochondrial Cytb gene of the rotifer Brachionus and a method for identifying the rotifer Brachionus. Background technique [0002] Rotifera (Rotifera) is a kind of small aquatic invertebrates that live in planktonic, benthic or periplexus. The general body length is between 50-2000um, and it belongs to the phylum Rotifera. Early rotifer studies focused on morphology and ultrastructure. Due to the variety of rotifer shapes and ecological habits, and the lack of in-depth understanding of its ultrastructure and biochemical properties, its evolutionary pathway has been difficult to determine. With the in-depth research on rotifers, the morphological data can no longer satisfy the identification and phylogenetic research of some species of rotifers. Contents of the invention [0003] Aiming at the deficiencies of the prior ar...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888
CPCC12Q1/6888C12Q2600/124
Inventor 穆维平杨宜冰王静夏月琴方锣泽张强程双怀
Owner ANHUI NORMAL UNIV
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