Method for partial rDNA (recombinant deoxyribonucleic acid) sequencing of mitochondrial 16S genes of Brachionus rotifer and identification of Brachionus rotifer

A technology of mitochondria and rotifers, applied in the field of biology

Inactive Publication Date: 2014-12-17
ANHUI NORMAL UNIV
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Problems solved by technology

The identification of rotifers by molecular biology solves the identification difficulties of using morphology and ultrastructure, so that in the process of large-scale cultivation of rotifers, the population competition caused by population mixing can be reduced, resulting in reduced production of rotifers or even possibility of mass death

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Embodiment Construction

[0014] The biological material used in this method for identifying the genus Brachionus rotifers is Brachionus angularis, Brachionus schizopoda, Brachiona redus, Brachiona square, Brachiona cucurbitus, Brachiona scissors Brachionus rotifers, Brachionus falciparum, Brachionus tendigitata, Brachionus rectangle, pick a single individual of each rotifer, and culture it as a single clone. Brachionus rotifer and Brachionus angularis adopt the formula of Gilbert (1963), and the other species use the original local water samples to filter and make the culture medium. The temperature is basically consistent with the ambient temperature at the time of collection. The food used is HB-4 For Scenedesmus obliquus in the exponential growth phase cultured in culture medium, when the number of individual cloned rotifers reaches a certain amount, the rotifers are filtered out with a 25 μm pore size filter, rinsed with sterile distilled water, and collected in Eppendof tubes , stored in a -20°C ...

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Abstract

The invention relates to a method for partial rDNA (recombinant deoxyribonucleic acid) sequencing of mitochondrial ND16S genes of Brachionus rotifer and identification of the Brachionus rotifer. The sequencing method comprises the following steps of: extracting total DNA (deoxyribonucleic acid) of the Brachionus rotifer, performing PCR (polymerase chain reaction) primer amplification, determining a sequence, and performing phylogenetic analysis. According to the identification method, mitochondrial ND16S rDNA of the Brachionus rotifer is used for identification. A designed PCR primer in the sequencing method can amplify the mitochondrial ND16S rDNA of Brachionus rubens and the like, a sequencing result can be used for precisely identifying the Brachionus rubens and the like, the difficulty in identification by utilizing morphology and an ultra-micro structure can be solved, and then the possibility of drop in production and even mass deaths of the rotifers caused by population competition in mixed populations during the large-scale culture of the rotifers can be further reduced.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a partial rDNA sequencing of the mitochondrial 16S gene of the rotifer of the genus Brachionus and a method for identifying the rotifer of the genus Brachionus. Background technique [0002] Early taxonomic and systematic studies of rotifers were mainly based on morphological data. For example, Wallace (1989) conducted cluster analysis using eight morphological features and found that Rotifera, Eutatoria, and Monogonota were monophyletic, and clustered using nine morphological features. Class analysis found that the genus Notholca was divided into six clades. Melone (1998) obtained a parsimony clustering tree through matrix analysis of 60 morphological features of 1 acanthocephala, 6 rotifers and 2 outgroups (planarians and gnathogastrics), and found that the Dinestidate ( Digononta) is a paraphyletic group, but the monophyletic of rotifers cannot be determined....

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 程双怀张逸夏梦宁刘应龙卢燕梅闫锦锦张宁
Owner ANHUI NORMAL UNIV
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